                         SEQUENCE LISTING

<110>  Danisco US Inc.
       Frisch, Ryan L.
       He, Hongxian L.
 
<120>  CAS9 VARIANTS AND METHODS OF USE

<130>  NB41317-PCT

<150>  US 62/599176
<151>  2017-12-15

<160>  157   

<170>  PatentIn version 3.5

<210>  1
<211>  1368
<212>  PRT
<213>  Streptococcus  pyogenes

<400>  1

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             
Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             
Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             
Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             
Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             
Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             
Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             
Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             
Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             
Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             
Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             
Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             
Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             
Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             
Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             
Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             
Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             
Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             
His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             
Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             
Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             
Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             
Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             
Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             
Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             

<210>  2
<211>  5056
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Bacillus codon optimized Cas9

<400>  2
aattcctcca ttttcttctg ctatcaaaat aacagactcg tgattttcca aacgagcttt       60
caaaaaagcc tctgcccctt gcaaatcgga tgcctgtcta taaaattccc gatattggtt      120
aaacagcggc gcaatggcgg ccgcatctga tgtctttgct tggcgaatgt tcatcttatt      180
tcttcctccc tctcaataat tttttcattc tatccctttt ctgtaaagtt tatttttcag      240
aatactttta tcatcatgct ttgaaaaaat atcacgataa tatccattgt tctcacggaa      300
gcacacgcag gtcatttgaa cgaatttttt cgacaggaat ttgccgggac tcaggagcat      360
ttaacctaaa aaagcatgac atttcagcat aatgaacatt tactcatgtc tattttcgtt      420
cttttctgta tgaaaatagt tatttcgagt ctctacggaa atagcgagag atgatatacc      480
taaatagaga taaaatcatc tcaaaaaaat gggtctacta aaatattatt ccatctatta      540
caataaattc acagaatagt cttttaagta agtctactct gaattttttt aaaaggagag      600
ggtaactagt ggccccaaaa aagaaacgca aggttatgga taaaaaatac agcattggtc      660
tggatatcgg aaccaacagc gttgggtggg cagtaataac agatgaatac aaagtgccgt      720
caaaaaaatt taaggttctg gggaatacag atcgccacag cataaaaaag aatctgattg      780
gggcattgct gtttgattcg ggtgagacag ctgaggccac gcgtctgaaa cgtacagcaa      840
gaagacgtta cacacgtcgt aaaaatcgta tttgctactt acaggaaatt ttttctaacg      900
aaatggccaa ggtagatgat agtttcttcc atcgtctcga agaatctttt ctggttgagg      960
aagataaaaa acacgaacgt caccctatct ttggcaatat cgtggatgaa gtggcctatc     1020
atgaaaaata ccctacgatt tatcatcttc gcaagaagtt ggttgatagt acggacaaag     1080
cggatctgcg tttaatctat cttgcgttag cgcacatgat caaatttcgt ggtcatttct     1140
taattgaagg tgatctgaat cctgataact ctgatgtgga caaattgttt atacaattag     1200
tgcaaaccta taatcagctg ttcgaggaaa accccattaa tgcctctgga gttgatgcca     1260
aagcgatttt aagcgcgaga ctttctaagt cccggcgtct ggagaatctg atcgcccagt     1320
taccagggga aaagaaaaat ggtctgtttg gtaatctgat tgccctcagt ctggggctta     1380
ccccgaactt caaatccaat tttgacctgg ctgaggacgc aaagctgcag ctgagcaaag     1440
atacttatga tgatgacctc gacaatctgc tcgcccagat tggtgaccaa tatgcggatc     1500
tgtttctggc agcgaagaat ctttcggatg ctatcttgct gtcggatatt ctgcgtgtta     1560
ataccgaaat caccaaagcg cctctgtctg caagtatgat caagagatac gacgagcacc     1620
accaggacct gactcttctt aaggcactgg tacgccaaca gcttccggag aaatacaaag     1680
aaatattctt cgaccagtcc aagaatggtt acgcgggcta catcgatggt ggtgcatcac     1740
aggaagagtt ctataaattt attaaaccaa tccttgagaa aatggatggc acggaagagt     1800
tacttgttaa acttaaccgc gaagacttgc ttagaaagca acgtacattc gacaacggct     1860
ccatcccaca ccagattcat ttaggtgaac ttcacgccat cttgcgcaga caagaagatt     1920
tctatccctt cttaaaagac aatcgggaga aaatcgagaa gatcctgacg ttccgcattc     1980
cctattatgt cggtcccctg gcacgtggta attctcggtt tgcctggatg acgcgcaaaa     2040
gtgaggaaac catcacccct tggaactttg aagaagtcgt ggataaaggt gctagcgcgc     2100
agtcttttat agaaagaatg acgaacttcg ataaaaactt gcccaacgaa aaagtcctgc     2160
ccaagcactc tcttttatat gagtacttta ctgtgtacaa cgaactgact aaagtgaaat     2220
acgttacgga aggtatgcgc aaacctgcct ttcttagtgg cgagcagaaa aaagcaattg     2280
tcgatcttct ctttaaaacg aatcgcaagg taactgtaaa acagctgaag gaagattatt     2340
tcaaaaagat cgaatgcttt gattctgtcg agatctcggg tgtcgaagat cgtttcaacg     2400
cttccttagg gacctatcat gatttgctga agataataaa agacaaagac tttctcgaca     2460
atgaagaaaa tgaagatatt ctggaggata ttgttttgac cttgacctta ttcgaagata     2520
gagagatgat cgaggagcgc ttaaaaacct atgcccacct gtttgatgac aaagtcatga     2580
agcaattaaa gcgccgcaga tatacggggt ggggccgctt gagccgcaag ttgattaacg     2640
gtattagaga caagcagagc ggaaaaacta tcctggattt cctcaaatct gacggatttg     2700
cgaaccgcaa ttttatgcag cttatacatg atgattcgct tacattcaaa gaggatattc     2760
agaaggctca ggtgtctggg caaggtgatt cactccacga acatatagca aatttggccg     2820
gctctcctgc gattaagaag gggatcctgc aaacagttaa agttgtggat gaacttgtaa     2880
aagtaatggg ccgccacaag ccggagaata tcgtgataga aatggcgcgc gagaatcaaa     2940
cgacacaaaa aggtcaaaag aactcaagag agagaatgaa gcgcattgag gaggggataa     3000
aggaacttgg atctcaaatt ctgaaagaac atccagttga aaacactcag ctgcaaaatg     3060
aaaaattgta cctgtactac ctgcagaatg gaagagacat gtacgtggat caggaattgg     3120
atatcaatag actctcggac tatgacgtag atcacattgt ccctcagagc ttcctcaagg     3180
atgattctat agataataaa gtacttacga gatcggacaa aaatcgcggt aaatcggata     3240
acgtcccatc ggaggaagtc gttaaaaaga tgaaaaacta ttggcgtcaa ctgctgaacg     3300
ccaagctgat cacacagcgt aagtttgata atctgactaa agccgaacgc ggtggtctta     3360
gtgaactcga taaagcagga tttataaaac ggcagttagt agaaacgcgc caaattacga     3420
aacacgtggc tcagatcctc gattctagaa tgaatacaaa gtacgatgaa aacgataaac     3480
tgatccgtga agtaaaagtc attaccttaa aatctaaact tgtgtccgat ttccgcaaag     3540
attttcagtt ttacaaggtc cgggaaatca ataactatca ccatgcacat gatgcatatt     3600
taaatgcggt tgtaggcacg gcccttatta agaaataccc taaactcgaa agtgagtttg     3660
tttatgggga ttataaagtg tatgacgttc gcaaaatgat cgcgaaatca gaacaggaaa     3720
tcggtaaggc taccgctaaa tacttttttt attccaacat tatgaatttt tttaagaccg     3780
aaataactct cgcgaatggt gaaatccgta aacggcctct tatagaaacc aatggtgaaa     3840
cgggagaaat cgtttgggat aaaggtcgtg actttgccac cgttcgtaaa gtcctctcaa     3900
tgccgcaagt taacattgtc aagaagacgg aagttcaaac agggggattc tccaaagaat     3960
ctatcctgcc gaagcgtaac agtgataaac ttattgccag aaaaaaagat tgggatccaa     4020
aaaaatacgg aggctttgat tcccctaccg tcgcgtatag tgtgctggtg gttgctaaag     4080
tcgagaaagg gaaaagcaag aaattgaaat cagttaaaga actgctgggt attacaatta     4140
tggaaagatc gtcctttgag aaaaatccga tcgacttttt agaggccaag gggtataagg     4200
aagtgaaaaa agatctcatc atcaaattac cgaagtatag tctttttgag ctggaaaacg     4260
gcagaaaaag aatgctggcc tccgcgggcg agttacagaa gggaaatgag ctggcgctgc     4320
cttccaaata tgttaatttt ctgtaccttg ccagtcatta tgagaaactg aagggcagcc     4380
ccgaagataa cgaacagaaa caattattcg tggaacagca taagcactat ttagatgaaa     4440
ttatagagca aattagtgaa ttttctaagc gcgttatcct cgcggatgct aatttagaca     4500
aagtactgtc agcttataat aaacatcggg ataagccgat tagagaacag gccgaaaata     4560
tcattcattt gtttacctta accaaccttg gagcaccagc tgccttcaaa tatttcgata     4620
ccacaattga tcgtaaacgg tatacaagta caaaagaagt cttggacgca accctcattc     4680
atcaatctat tactggatta tatgagacac gcattgatct ttcacagctg ggcggagaca     4740
agaagaaaaa actgaaactg caccatcatc accatcatca tcaccatcat tgataactcg     4800
agaaagctta cataaaaaac cggccttggc cccgccggtt ttttattatt tttcttcctc     4860
cgcatgttca atccgctcca taatcgacgg atggctccct ctgaaaattt taacgagaaa     4920
cggcgggttg acccggctca gtcccgtaac ggccaagtcc tgaaacgtct caatcgccgc     4980
ttcccggttt ccggtcagct caatgccgta acggtcggcg gcgttttcct gataccggga     5040
gacggcattc gtaatc                                                     5056

<210>  3
<211>  8
<212>  PRT
<213>  Artificial sequence

<220>
<223>  N-terminal NLS

<400>  3
Ala Pro Lys Lys Lys Arg Lys Val 
1               5               

<210>  4
<211>  6
<212>  PRT
<213>  Artificial sequence

<220>
<223>  C-terminal NLS

<400>  4
Lys Lys Lys Lys Leu Lys 
1               5       

<210>  5
<211>  10
<212>  PRT
<213>  Artificial sequence

<220>
<223>  deca-Histidine tag

<400>  5
His His His His His His His His His His 
1               5                   10  

<210>  6
<211>  607
<212>  DNA
<213>  Bacillus subtilis

<400>  6
attcctccat tttcttctgc tatcaaaata acagactcgt gattttccaa acgagctttc       60
aaaaaagcct ctgccccttg caaatcggat gcctgtctat aaaattcccg atattggtta      120
aacagcggcg caatggcggc cgcatctgat gtctttgctt ggcgaatgtt catcttattt      180
cttcctccct ctcaataatt ttttcattct atcccttttc tgtaaagttt atttttcaga      240
atacttttat catcatgctt tgaaaaaata tcacgataat atccattgtt ctcacggaag      300
cacacgcagg tcatttgaac gaattttttc gacaggaatt tgccgggact caggagcatt      360
taacctaaaa aagcatgaca tttcagcata atgaacattt actcatgtct attttcgttc      420
ttttctgtat gaaaatagtt atttcgagtc tctacggaaa tagcgagaga tgatatacct      480
aaatagagat aaaatcatct caaaaaaatg ggtctactaa aatattattc catctattac      540
aataaattca cagaatagtc ttttaagtaa gtctactctg aattttttta aaaggagagg      600
gtaacta                                                                607

<210>  7
<211>  247
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized terminator DNA sequence

<400>  7
acataaaaaa ccggccttgg ccccgccggt tttttattat ttttcttcct ccgcatgttc       60
aatccgctcc ataatcgacg gatggctccc tctgaaaatt ttaacgagaa acggcgggtt      120
gacccggctc agtcccgtaa cggccaagtc ctgaaacgtc tcaatcgccg cttcccggtt      180
tccggtcagc tcaatgccgt aacggtcggc ggcgttttcc tgataccggg agacggcatt      240
cgtaatc                                                                247

<210>  8
<211>  50
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cas9 forward primer

<400>  8
atatatgagt aaacttggtc tgacagaatt cctccatttt cttctgctat                  50

<210>  9
<211>  35
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cas9 revers primer

<400>  9
tgcggccgcg aattcgatta cgaatgccgt ctccc                                  35

<210>  10
<211>  3290
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pKB320 backbone

<400>  10
gaattcgcgg ccgcacgcgt ccatggggat ccccgcgggt cgacctcgag agttacgcta       60
gggataacag ggtaatatag gagctccagt cggcttaaac cagttttcgc tggtgcgaaa      120
aaagagtgtc ttgtgacacc taaattcaaa atctatcggt cagatttata ccgatttgat      180
tttatatatt cttgaataac atacgccgag ttatcacata aaagcgggaa ccaatcataa      240
aatttaaact tcattgcata atccattaaa ctcttaaatt ctacgattcc ttgttcatca      300
ataaactcaa tcatttcttt aattaattta tatctatctg ttgttgtttt ctttaataat      360
tcattaacat ctacaccgcc ataaactatc atatcttctt tttgatattt aaatttatta      420
ggatcgtcca tgtgaagcat atatctcaca agacctttca cacttcctgc aatctgcgga      480
atagtcgcat tcaattcttc tgttaattat ttttatctgt tcataagatt tattaccctc      540
atacatcact agaatatgat aatgctcttt tttcatccta ccttctgtat cagtatccct      600
atcatgtaat ggagacacta caaattgaat gtgtaactct tttaaatact ctaaccactc      660
ggcttttgct gattctggat ataaaacaaa tgtccaatta cgtcctcttg aatttttctt      720
gttttcagtt tcttttatta cattttcgct catgatataa taacggtgct aatacactta      780
acaaaattta gtcatagata ggcagcatgc cagtgctgtc tatctttttt tgtttaaaat      840
gcaccgtatt cctcctttgc atattttttt attagaatac cggttgcatc tgatttgcta      900
atattatatt tttctttgat tctatttaat atctcatttt cttctgttgt aagtcttaaa      960
gtaacagcaa cttttttctc ttcttttcta tctacaacta tcactgtacc tcccaacatc     1020
tgtttttttc actttaacat aaaaaacaac cttttaacat taaaaaccca atatttattt     1080
atttgtttgg acaatggaca ctggacacct aggggggagg tcgtagtacc cccctatgtt     1140
ttctccccta aataacccca aaaatctaag aaaaaaagac ctcaaaaagg tctttaatta     1200
acatctcaaa tttcgcattt attccaattt cctttttgcg tgtgatgcga gctcatcggc     1260
tccgtcgata ctatgttata cgccaacttt caaaacaact ttgaaaaagc tgttttctgg     1320
tatttaaggt tttagaatgc aaggaacagt gaattggagt tcgtcttgtt ataattagct     1380
tcttggggta tctttaaata ctgtagaaaa gaggaaggaa ataataaatg gctaaaatga     1440
gaatatcacc ggaattgaaa aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag     1500
gaatgtctcc tgctaaggta tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa     1560
tgacggacag ccggtataaa gggaccacct atgatgtgga acgggaaaag gacatgatgc     1620
tatggctgga aggaaagctg cctgttccaa aggtcctgca ctttgaacgg catgatggct     1680
ggagcaatct gctcatgagt gaggccgatg gcgtcctttg ctcggaagag tatgaagatg     1740
aacaaagccc tgaaaagatt atcgagctgt atgcggagtg catcaggctc tttcactcca     1800
tcgacatatc ggattgtccc tatacgaata gcttagacag ccgcttagcc gaattggatt     1860
acttactgaa taacgatctg gccgatgtgg attgcgaaaa ctgggaagaa gacactccat     1920
ttaaagatcc gcgcgagctg tatgattttt taaagacgga aaagcccgaa gaggaacttg     1980
tcttttccca cggcgacctg ggagacagca acatctttgt gaaagatggc aaagtaagtg     2040
gctttattga tcttgggaga agcggcaggg cggacaagtg gtatgacatt gccttctgcg     2100
tccggtcgat cagggaggat atcggggaag aacagtatgt cgagctattt tttgacttac     2160
tggggatcaa gcctgattgg gagaaaataa aatattatat tttactggat gaattgtttt     2220
agtgactgca gtgagatctg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag     2280
gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg     2340
agtaggacaa atccgccgct ctagctaagc agaaggccat cctgacggat ggcctttttg     2400
cgtttctaca aactcttgtt aactctagag ctgcctgccg cgtttcggtg atgaagatct     2460
tcccgatgat taattaattc agaacgctcg gttgccgccg ggcgtttttt atgaagcttc     2520
gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc     2580
aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag     2640
ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct     2700
cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta     2760
ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc     2820
cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc     2880
agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt     2940
gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct     3000
gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc     3060
tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca     3120
agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta     3180
agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa     3240
atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca                3290

<210>  11
<211>  4204
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pKB320 plasmid

<400>  11
gcggccgcac gcgtccatgg ggatccccgc gggtcgacct cgagagttac gctagggata       60
acagggtaat ataggagctc cagtcggctt aaaccagttt tcgctggtgc gaaaaaagag      120
tgtcttgtga cacctaaatt caaaatctat cggtcagatt tataccgatt tgattttata      180
tattcttgaa taacatacgc cgagttatca cataaaagcg ggaaccaatc ataaaattta      240
aacttcattg cataatccat taaactctta aattctacga ttccttgttc atcaataaac      300
tcaatcattt ctttaattaa tttatatcta tctgttgttg ttttctttaa taattcatta      360
acatctacac cgccataaac tatcatatct tctttttgat atttaaattt attaggatcg      420
tccatgtgaa gcatatatct cacaagacct ttcacacttc ctgcaatctg cggaatagtc      480
gcattcaatt cttctgttaa ttatttttat ctgttcataa gatttattac cctcatacat      540
cactagaata tgataatgct cttttttcat cctaccttct gtatcagtat ccctatcatg      600
taatggagac actacaaatt gaatgtgtaa ctcttttaaa tactctaacc actcggcttt      660
tgctgattct ggatataaaa caaatgtcca attacgtcct cttgaatttt tcttgttttc      720
agtttctttt attacatttt cgctcatgat ataataacgg tgctaataca cttaacaaaa      780
tttagtcata gataggcagc atgccagtgc tgtctatctt tttttgttta aaatgcaccg      840
tattcctcct ttgcatattt ttttattaga ataccggttg catctgattt gctaatatta      900
tatttttctt tgattctatt taatatctca ttttcttctg ttgtaagtct taaagtaaca      960
gcaacttttt tctcttcttt tctatctaca actatcactg tacctcccaa catctgtttt     1020
tttcacttta acataaaaaa caacctttta acattaaaaa cccaatattt atttatttgt     1080
ttggacaatg gacactggac acctaggggg gaggtcgtag taccccccta tgttttctcc     1140
cctaaataac cccaaaaatc taagaaaaaa agacctcaaa aaggtcttta attaacatct     1200
caaatttcgc atttattcca atttcctttt tgcgtgtgat gcgagctcat cggctccgtc     1260
gatactatgt tatacgccaa ctttcaaaac aactttgaaa aagctgtttt ctggtattta     1320
aggttttaga atgcaaggaa cagtgaattg gagttcgtct tgttataatt agcttcttgg     1380
ggtatcttta aatactgtag aaaagaggaa ggaaataata aatggctaaa atgagaatat     1440
caccggaatt gaaaaaactg atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt     1500
ctcctgctaa ggtatataag ctggtgggag aaaatgaaaa cctatattta aaaatgacgg     1560
acagccggta taaagggacc acctatgatg tggaacggga aaaggacatg atgctatggc     1620
tggaaggaaa gctgcctgtt ccaaaggtcc tgcactttga acggcatgat ggctggagca     1680
atctgctcat gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa     1740
gccctgaaaa gattatcgag ctgtatgcgg agtgcatcag gctctttcac tccatcgaca     1800
tatcggattg tccctatacg aatagcttag acagccgctt agccgaattg gattacttac     1860
tgaataacga tctggccgat gtggattgcg aaaactggga agaagacact ccatttaaag     1920
atccgcgcga gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt     1980
cccacggcga cctgggagac agcaacatct ttgtgaaaga tggcaaagta agtggcttta     2040
ttgatcttgg gagaagcggc agggcggaca agtggtatga cattgccttc tgcgtccggt     2100
cgatcaggga ggatatcggg gaagaacagt atgtcgagct attttttgac ttactgggga     2160
tcaagcctga ttgggagaaa ataaaatatt atattttact ggatgaattg ttttagtgac     2220
tgcagtgaga tctggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag     2280
tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg     2340
acaaatccgc cgctctagct aagcagaagg ccatcctgac ggatggcctt tttgcgtttc     2400
tacaaactct tgttaactct agagctgcct gccgcgtttc ggtgatgaag atcttcccga     2460
tgattaatta attcagaacg ctcggttgcc gccgggcgtt ttttatgaag cttcgttgct     2520
ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca     2580
gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct     2640
cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc     2700
gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt     2760
tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc     2820
cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc     2880
cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg     2940
gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc     3000
agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag     3060
cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga     3120
tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat     3180
tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag     3240
ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat     3300
cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc     3360
cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat     3420
accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag     3480
ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg     3540
ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc     3600
tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca     3660
acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg     3720
tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc     3780
actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta     3840
ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc     3900
aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg     3960
ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc     4020
cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc     4080
aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat     4140
actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgga     4200
attc                                                                  4204

<210>  12
<211>  35
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pKB320 forward primer

<400>  12
gggagacggc attcgtaatc gaattcgcgg ccgca                                  35

<210>  13
<211>  50
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pKB320 reverse primer

<400>  13
atagcagaag aaaatggagg aattctgtca gaccaagttt actcatatat                  50

<210>  14
<211>  23
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid RSP1

<400>  14
ccgactggag ctcctatatt acc                                               23

<210>  15
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid RSP2

<400>  15
gctgtggcga tctgtattcc                                                   20

<210>  16
<211>  22
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP1

<400>  16
gtcttttaag taagtctact ct                                                22

<210>  17
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP2

<400>  17
ccaaagcgat tttaagcgcg                                                   20

<210>  18
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP3

<400>  18
cctggcacgt ggtaattctc                                                   20

<210>  19
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP4

<400>  19
ggatttcctc aaatctgacg                                                   20

<210>  20
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP5

<400>  20
gtagaaacgc gccaaattac g                                                 21

<210>  21
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP6

<400>  21
gctggtggtt gctaaagtcg                                                   20

<210>  22
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP7

<400>  22
ggacgcaacc ctcattcatc                                                   20

<210>  23
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid RSP3

<400>  23
caggcatccg atttgcaagg                                                   20

<210>  24
<211>  19
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid FSP8

<400>  24
gcaagcagca gattacgcg                                                    19

<210>  25
<211>  8347
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF694

<400>  25
gaattcctcc attttcttct gctatcaaaa taacagactc gtgattttcc aaacgagctt       60
tcaaaaaagc ctctgcccct tgcaaatcgg atgcctgtct ataaaattcc cgatattggt      120
taaacagcgg cgcaatggcg gccgcatctg atgtctttgc ttggcgaatg ttcatcttat      180
ttcttcctcc ctctcaataa ttttttcatt ctatcccttt tctgtaaagt ttatttttca      240
gaatactttt atcatcatgc tttgaaaaaa tatcacgata atatccattg ttctcacgga      300
agcacacgca ggtcatttga acgaattttt tcgacaggaa tttgccggga ctcaggagca      360
tttaacctaa aaaagcatga catttcagca taatgaacat ttactcatgt ctattttcgt      420
tcttttctgt atgaaaatag ttatttcgag tctctacgga aatagcgaga gatgatatac      480
ctaaatagag ataaaatcat ctcaaaaaaa tgggtctact aaaatattat tccatctatt      540
acaataaatt cacagaatag tcttttaagt aagtctactc tgaatttttt taaaaggaga      600
gggtaactag tggccccaaa aaagaaacgc aaggttatgg ataaaaaata cagcattggt      660
ctggatatcg gaaccaacag cgttgggtgg gcagtaataa cagatgaata caaagtgccg      720
tcaaaaaaat ttaaggttct ggggaataca gatcgccaca gcataaaaaa gaatctgatt      780
ggggcattgc tgtttgattc gggtgagaca gctgaggcca cgcgtctgaa acgtacagca      840
agaagacgtt acacacgtcg taaaaatcgt atttgctact tacaggaaat tttttctaac      900
gaaatggcca aggtagatga tagtttcttc catcgtctcg aagaatcttt tctggttgag      960
gaagataaaa aacacgaacg tcaccctatc tttggcaata tcgtggatga agtggcctat     1020
catgaaaaat accctacgat ttatcatctt cgcaagaagt tggttgatag tacggacaaa     1080
gcggatctgc gtttaatcta tcttgcgtta gcgcacatga tcaaatttcg tggtcatttc     1140
ttaattgaag gtgatctgaa tcctgataac tctgatgtgg acaaattgtt tatacaatta     1200
gtgcaaacct ataatcagct gttcgaggaa aaccccatta atgcctctgg agttgatgcc     1260
aaagcgattt taagcgcgag actttctaag tcccggcgtc tggagaatct gatcgcccag     1320
ttaccagggg aaaagaaaaa tggtctgttt ggtaatctga ttgccctcag tctggggctt     1380
accccgaact tcaaatccaa ttttgacctg gctgaggacg caaagctgca gctgagcaaa     1440
gatacttatg atgatgacct cgacaatctg ctcgcccaga ttggtgacca atatgcggat     1500
ctgtttctgg cagcgaagaa tctttcggat gctatcttgc tgtcggatat tctgcgtgtt     1560
aataccgaaa tcaccaaagc gcctctgtct gcaagtatga tcaagagata cgacgagcac     1620
caccaggacc tgactcttct taaggcactg gtacgccaac agcttccgga gaaatacaaa     1680
gaaatattct tcgaccagtc caagaatggt tacgcgggct acatcgatgg tggtgcatca     1740
caggaagagt tctataaatt tattaaacca atccttgaga aaatggatgg cacggaagag     1800
ttacttgtta aacttaaccg cgaagacttg cttagaaagc aacgtacatt cgacaacggc     1860
tccatcccac accagattca tttaggtgaa cttcacgcca tcttgcgcag acaagaagat     1920
ttctatccct tcttaaaaga caatcgggag aaaatcgaga agatcctgac gttccgcatt     1980
ccctattatg tcggtcccct ggcacgtggt aattctcggt ttgcctggat gacgcgcaaa     2040
agtgaggaaa ccatcacccc ttggaacttt gaagaagtcg tggataaagg tgctagcgcg     2100
cagtctttta tagaaagaat gacgaacttc gataaaaact tgcccaacga aaaagtcctg     2160
cccaagcact ctcttttata tgagtacttt actgtgtaca acgaactgac taaagtgaaa     2220
tacgttacgg aaggtatgcg caaacctgcc tttcttagtg gcgagcagaa aaaagcaatt     2280
gtcgatcttc tctttaaaac gaatcgcaag gtaactgtaa aacagctgaa ggaagattat     2340
ttcaaaaaga tcgaatgctt tgattctgtc gagatctcgg gtgtcgaaga tcgtttcaac     2400
gcttccttag ggacctatca tgatttgctg aagataataa aagacaaaga ctttctcgac     2460
aatgaagaaa atgaagatat tctggaggat attgttttga ccttgacctt attcgaagat     2520
agagagatga tcgaggagcg cttaaaaacc tatgcccacc tgtttgatga caaagtcatg     2580
aagcaattaa agcgccgcag atatacgggg tggggccgct tgagccgcaa gttgattaac     2640
ggtattagag acaagcagag cggaaaaact atcctggatt tcctcaaatc tgacggattt     2700
gcgaaccgca attttatgca gcttatacat gatgattcgc ttacattcaa agaggatatt     2760
cagaaggctc aggtgtctgg gcaaggtgat tcactccacg aacatatagc aaatttggcc     2820
ggctctcctg cgattaagaa ggggatcctg caaacagtta aagttgtgga tgaacttgta     2880
aaagtaatgg gccgccacaa gccggagaat atcgtgatag aaatggcgcg cgagaatcaa     2940
acgacacaaa aaggtcaaaa gaactcaaga gagagaatga agcgcattga ggaggggata     3000
aaggaacttg gatctcaaat tctgaaagaa catccagttg aaaacactca gctgcaaaat     3060
gaaaaattgt acctgtacta cctgcagaat ggaagagaca tgtacgtgga tcaggaattg     3120
gatatcaata gactctcgga ctatgacgta gatcacattg tccctcagag cttcctcaag     3180
gatgattcta tagataataa agtacttacg agatcggaca aaaatcgcgg taaatcggat     3240
aacgtcccat cggaggaagt cgttaaaaag atgaaaaact attggcgtca actgctgaac     3300
gccaagctga tcacacagcg taagtttgat aatctgacta aagccgaacg cggtggtctt     3360
agtgaactcg ataaagcagg atttataaaa cggcagttag tagaaacgcg ccaaattacg     3420
aaacacgtgg ctcagatcct cgattctaga atgaatacaa agtacgatga aaacgataaa     3480
ctgatccgtg aagtaaaagt cattacctta aaatctaaac ttgtgtccga tttccgcaaa     3540
gattttcagt tttacaaggt ccgggaaatc aataactatc accatgcaca tgatgcatat     3600
ttaaatgcgg ttgtaggcac ggcccttatt aagaaatacc ctaaactcga aagtgagttt     3660
gtttatgggg attataaagt gtatgacgtt cgcaaaatga tcgcgaaatc agaacaggaa     3720
atcggtaagg ctaccgctaa atactttttt tattccaaca ttatgaattt ttttaagacc     3780
gaaataactc tcgcgaatgg tgaaatccgt aaacggcctc ttatagaaac caatggtgaa     3840
acgggagaaa tcgtttggga taaaggtcgt gactttgcca ccgttcgtaa agtcctctca     3900
atgccgcaag ttaacattgt caagaagacg gaagttcaaa cagggggatt ctccaaagaa     3960
tctatcctgc cgaagcgtaa cagtgataaa cttattgcca gaaaaaaaga ttgggatcca     4020
aaaaaatacg gaggctttga ttcccctacc gtcgcgtata gtgtgctggt ggttgctaaa     4080
gtcgagaaag ggaaaagcaa gaaattgaaa tcagttaaag aactgctggg tattacaatt     4140
atggaaagat cgtcctttga gaaaaatccg atcgactttt tagaggccaa ggggtataag     4200
gaagtgaaaa aagatctcat catcaaatta ccgaagtata gtctttttga gctggaaaac     4260
ggcagaaaaa gaatgctggc ctccgcgggc gagttacaga agggaaatga gctggcgctg     4320
ccttccaaat atgttaattt tctgtacctt gccagtcatt atgagaaact gaagggcagc     4380
cccgaagata acgaacagaa acaattattc gtggaacagc ataagcacta tttagatgaa     4440
attatagagc aaattagtga attttctaag cgcgttatcc tcgcggatgc taatttagac     4500
aaagtactgt cagcttataa taaacatcgg gataagccga ttagagaaca ggccgaaaat     4560
atcattcatt tgtttacctt aaccaacctt ggagcaccag ctgccttcaa atatttcgat     4620
accacaattg atcgtaaacg gtatacaagt acaaaagaag tcttggacgc aaccctcatt     4680
catcaatcta ttactggatt atatgagaca cgcattgatc tttcacagct gggcggagac     4740
aagaagaaaa aactgaaact gcaccatcat caccatcatc atcaccatca ttgataactc     4800
gagaaagctt acataaaaaa ccggccttgg ccccgccggt tttttattat ttttcttcct     4860
ccgcatgttc aatccgctcc ataatcgacg gatggctccc tctgaaaatt ttaacgagaa     4920
acggcgggtt gacccggctc agtcccgtaa cggccaagtc ctgaaacgtc tcaatcgccg     4980
cttcccggtt tccggtcagc tcaatgccgt aacggtcggc ggcgttttcc tgataccggg     5040
agacggcatt cgtaatcgaa ttcgcggccg cacgcgtcca tggggatccc cgcgggtcga     5100
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag     5160
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag     5220
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa     5280
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta     5340
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg     5400
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt     5460
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac     5520
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca     5580
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct     5640
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt     5700
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt     5760
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa     5820
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat     5880
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg     5940
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt     6000
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca     6060
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa     6120
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg     6180
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc     6240
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt     6300
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg     6360
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg     6420
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata     6480
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg     6540
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga     6600
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg     6660
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt     6720
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc     6780
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat     6840
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg     6900
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg     6960
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa     7020
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa     7080
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta     7140
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga     7200
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt     7260
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc     7320
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt     7380
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct     7440
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt     7500
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc     7560
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat     7620
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag     7680
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga     7740
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg     7800
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt     7860
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac     7920
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc     7980
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt     8040
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc     8100
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc     8160
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg     8220
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag     8280
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg     8340
tctgaca                                                               8347

<210>  26
<211>  9724
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF801

<400>  26
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca      240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga      420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag      480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag      540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa      600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta      660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg      720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt      780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac      840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca      900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct      960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt     1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt     1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa     1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat     1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg     1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt     1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca     1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa     1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg     1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc     1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt     1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg     1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg     1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata     1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg     1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga     1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg     1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt     2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc     2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat     2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg     2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg     2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa     2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa     2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta     2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga     2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt     2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc     2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt     2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct     2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt     2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc     2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat     2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag     3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga     3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg     3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt     3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac     3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc     3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt     3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc     3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc     3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg     3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag     3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg     3660
tctgacaaat ggttctttcc cctgtcctaa acaaaaaacc cgctttattg aaaaagcggg     3720
gctgttttac agacaggtca aataaacgtt tgaaaatgtt catttcaaaa cgcgcggaac     3780
ctccatcttc tcccatccag actatactgt cggcttcgga atcgcaccga atcctgccca     3840
taaaaaggct cgcgggctta gagcgcttgc tcatcaccgc cggtagggaa tttcaccctg     3900
ccccgaagat tgatcttatt tatttttaat actgatatta ttataaatta attgtgaaaa     3960
aatgtacagg tgcaaagctt attgcgctgt tttgggacat cctgcacgat atttcggtaa     4020
actcactttt tccgcatact aaaaaccgca cattcacagt tatttcattt ttaattttcg     4080
tctttccgcg tgaaactcat tgacactctt tatggaatat ggtaaattat cagatattta     4140
tgacgcttat ttaggaggaa atcttacaca gaagctgcgg aacctgaaaa gaattccttt     4200
caggttccgt tttttttagg aattctccct gatctcaagc atctggcggg gataaatccg     4260
ctctcctttc aaatcgttcc attctttgag gcgctgtaca gttacgccca ttttttcggc     4320
gatatgatga agcgtatccc ctttccgcac tacatatgta ccggtcttcg attcatcgtc     4380
atgaaggcgg agtgtttggc cggccttgag atttgaatgt ttcaacccgt ttattctcat     4440
gatctcctcg atggatatac cgctatcctt gctgattctc cagagcgtgt cccctttttg     4500
aacggtcacc gcaccgctca ttgtcccggc gttttgataa acgtggatag aattttgccg     4560
gaacgcctcc tcacgaagca ccgtcagcgg attgattgca tatcttttat cttcagtcca     4620
tgaaccgtga tgcatttcaa aatgcaggtg ggttccggtc gatattcgaa ttcctccatt     4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc     4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc     4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc     4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc     4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt     4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa     5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg     5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata     5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac     5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg     5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa     5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta     5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt     5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca     5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg     5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac     5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc     5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt     5760
taatctatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg     5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata     5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa     5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa     6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca     6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg     6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag     6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca     6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga     6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg     6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct     6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac     6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc     6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct     6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg     6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca     6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag     6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc     6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag     6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct     6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg     7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga     7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg     7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg     7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc     7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca     7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt     7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg     7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga     7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc     7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag     7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat     7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc     7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac     7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag     7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg     7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca     7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata     8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc     8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag     8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt     8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg     8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt     8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta     8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg     8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg     8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta     8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga     8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag     8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga     8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt     8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag     8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa     8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg     9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg     9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa     9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag     9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt     9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc     9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta     9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac     9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca     9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat     9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac     9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc     9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt     9720
aatc                                                                  9724

<210>  27
<211>  9724
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF806 plasmid

<400>  27
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagatgcc atcagttcct      240
catagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga      420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag      480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag      540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa      600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta      660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg      720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt      780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac      840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca      900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct      960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt     1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt     1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa     1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat     1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg     1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt     1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca     1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa     1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg     1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc     1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt     1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg     1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg     1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata     1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg     1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga     1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg     1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt     2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc     2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat     2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg     2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg     2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa     2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa     2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta     2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga     2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt     2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc     2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt     2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct     2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt     2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc     2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat     2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag     3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga     3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg     3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt     3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac     3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc     3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt     3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc     3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc     3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg     3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag     3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg     3660
tctgacattg atattcagca ccctgcgcat ttcgaccggg agaacgactc tgccgagctc     3720
atcgattctc cggacaatcc cggtattttt cacgtttgaa aagcctcctt ttctcctttc     3780
tttattgact tttgtcaaca tctttataat aaaagagatc ttcaaatttt ttgttgaaat     3840
actgaatcat ctttccgatc acaagttgtc cgggcctcct ttcgccattt aaaactctgc     3900
tgagtgtcgc cggggatacg ccgatttcaa tggcaagctg atttaaggag agattgtgtt     3960
caatcatgta ctggagaaca aaatctcttt tgatatgaat cttttttacc atgattactc     4020
ccctttctaa tctcttatgt ttctttttat ctacattgaa catatacgat ttgttaactt     4080
ttgtcaatac ttttaccatc catatgtttc ctataggcaa tattcgtact aaaatatttt     4140
ataataagag attgcgaggt tttggccata cttctccgcg gcacactctc ctctctatca     4200
ttttcgtctg tttacgatcc tgctgttatt ttatccctta tgttaacttt tgtcaatatt     4260
tttcctgtct aagtatttcc tatagtcaac atttgtatta aaatgttcat atcatgaatt     4320
tgcggggggg atggcgatga caaggttcgg cgagcggctc aaagagctga gggaacaaag     4380
aagcctgtcg gttaatcagc ttgccatgta tgccggtgtg agcgccgcag ccatttccag     4440
agccgcagcc atttccagaa tcgaaaacgg ccaccgcggc gttcccaagc ccgcgacgat     4500
cagaaaattg gccgaggctc tgaaaatgcc gtacgagcag ctcatggata ttgccggtta     4560
tatgagagct gacgagattc gcgaacagcc gcgcggctat gtcacgatgc aggagatcgc     4620
ggccaagcac ggcgtcgaag acctgtggct gtttaaaccc gagaaatgaa ttcctccatt     4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc     4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc     4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc     4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc     4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt     4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa     5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg     5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata     5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac     5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg     5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa     5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta     5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt     5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca     5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg     5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac     5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc     5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt     5760
taatctatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg     5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata     5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa     5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa     6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca     6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg     6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag     6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca     6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga     6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg     6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct     6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac     6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc     6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct     6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg     6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca     6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag     6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc     6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag     6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct     6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg     7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga     7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg     7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg     7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc     7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca     7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt     7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg     7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga     7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc     7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag     7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat     7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc     7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac     7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag     7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg     7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca     7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata     8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc     8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag     8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt     8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg     8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt     8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta     8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg     8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg     8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta     8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga     8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag     8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga     8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt     8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag     8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa     8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg     9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg     9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa     9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag     9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt     9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc     9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta     9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac     9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca     9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat     9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac     9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc     9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt     9720
aatc                                                                  9724

<210>  28
<211>  20
<212>  DNA
<213>  Bacillus licheniformis

<400>  28
ctcgacttcg aatacatcca                                                   20

<210>  29
<211>  20
<212>  DNA
<213>  Bacillus licheniformis

<400>  29
gatgccatca gttcctcata                                                   20

<210>  30
<211>  1578
<212>  DNA
<213>  Bacillus licheniformis

<400>  30
atgtttcgag tattggtctc agataaaatg tccagcgacg gcctcaaacc attaatggaa       60
gcagatttta ttgaaattgt agaaaagaat gttgcggaag cggaagacga gcttcatacg      120
tttgacgcgc tcttggtgcg gagcgccacg aaggtaaccg aagagctgtt taaaaagatg      180
acttcgctga aaatcgtcgc cagagcaggt gtcggcgtcg acaatatcga tattgacgag      240
gcgacaaaac acggtgttat cgtcgtaaac gcgccaaacg ggaatacaat ttcaaccgct      300
gaacatacct ttgcaatgtt ttcagcgtta atgagacata ttccgcaggc aaacatctcc      360
gtgaaatcaa gggagtggaa tcgttcggct tacgtcggtt cagagcttta cggaaaaacg      420
ctcggcatca tcggaatggg ccgcatcgga agcgaaatcg cgagccgcgc aaaagcattc      480
ggtatgaccg ttcatgtatt tgacccgttc ctgacccaag aaagggcaag caagctcggc      540
gttaacgcga acagctttga agaagttctg gcatgcgccg acatcattac ggttcatacc      600
ccgctcacga aagaaacgaa gggacttttg aacaaagaaa ccatcgcaaa aacgaaaaaa      660
ggcgttcgtc tcgttaactg tgcaagaggc ggcatcatcg atgaagcagc gcttttggaa      720
gctctggaaa gcggacatgt cgctggcgct gccttggatg tattcgaagt cgagcctccg      780
gtcgattcaa aactgatcga tcatccgctt gtagtcgcga ctcctcactt gggcgcctca      840
acaaaagaag cccagctgaa tgtcgctgca caagtgtccg aagaagtcct tcagtatgcg      900
caaggaaacc ctgtgatgtc cgcgatcaac cttccggcca tgacaaagga ttcattcgaa      960
aaaatccagc cttatcatca gtttgccaat acgatcggaa accttgtgtc tcagtgcatg     1020
aatgagcctg ttcaagatgt agccatccaa tatgaaggct ccatcgccaa acttgaaacg     1080
tcatttatta cgaaaagcct tttggccgga tttctgaagc cgagggtcgc ggctaccgtt     1140
aacgaagtga atgccggcac cgttgcgaaa gagcgcggca tcagcttcag cgaaaaaatt     1200
tcttccaatg agtcaggcta tgaaaactgc atctctgtga ctgtcacggg agatgtaaca     1260
acattctctt taagagcgac gtacattccg cacttcggcg gacgcatcgt tgccttaaac     1320
ggctttgata ttgattttta tccggctgga caccttgtct acattcacca ccaggataaa     1380
ccaggggcta tcggccatgt cggacgaatt ttaggagacc atgacatcaa tatcgccact     1440
atgcaggtag gccgaaaaga aaaaggcgga gaagcgatca tgatgctttc ctttgaccgc     1500
caccttgagg acgatatttt agctgagctg aaaaacatcc cggatatcgt gtctgttaaa     1560
gccatcgacc ttccttaa                                                   1578

<210>  31
<211>  23
<212>  DNA
<213>  Bacillus licheniformis

<400>  31
ctcgacttcg aatacatcca agg                                               23

<210>  32
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized DNA encoding variable targeting domain

<400>  32
ctcgacttcg aatacatcca                                                   20

<210>  33
<211>  76
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized DNA encoding CER domain

<400>  33
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60
ggcaccgagt cggtgc                                                       76

<210>  34
<211>  96
<212>  RNA
<213>  Artificial sequence

<220>
<223>  synthesized gRNA targeting target site 1

<400>  34
cucgacuucg aauacaucca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60
cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96

<210>  35
<211>  224
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthesized spac promoter

<400>  35
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tgga                       224

<210>  36
<211>  95
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized  t0 terminator

<400>  36
gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc       60
tcggttgccg ccgggcgttt tttattggtg agaat                                  95

<210>  37
<211>  500
<212>  DNA
<213>  Bacillus licheniformis

<400>  37
aatggttctt tcccctgtcc taaacaaaaa acccgcttta ttgaaaaagc ggggctgttt       60
tacagacagg tcaaataaac gtttgaaaat gttcatttca aaacgcgcgg aacctccatc      120
ttctcccatc cagactatac tgtcggcttc ggaatcgcac cgaatcctgc ccataaaaag      180
gctcgcgggc ttagagcgct tgctcatcac cgccggtagg gaatttcacc ctgccccgaa      240
gattgatctt atttattttt aatactgata ttattataaa ttaattgtga aaaaatgtac      300
aggtgcaaag cttattgcgc tgttttggga catcctgcac gatatttcgg taaactcact      360
ttttccgcat actaaaaacc gcacattcac agttatttca tttttaattt tcgtctttcc      420
gcgtgaaact cattgacact ctttatggaa tatggtaaat tatcagatat ttatgacgct      480
tatttaggag gaaatcttac                                                  500

<210>  38
<211>  40
<212>  DNA
<213>  Artificial sequence

<220>
<223>  serA1 homology arm 1 forward primer

<400>  38
tgagtaaact tggtctgaca aatggttctt tcccctgtcc                             40

<210>  39
<211>  46
<212>  DNA
<213>  Artificial sequence

<220>
<223>  serA1 homology arm 1 forward primer

<400>  39
aggttccgca gcttctgtgt aagatttcct cctaaataag cgtcat                      46

<210>  40
<211>  500
<212>  DNA
<213>  Bacillus  licheniformis

<400>  40
acagaagctg cggaacctga aaagaattcc tttcaggttc cgtttttttt aggaattctc       60
cctgatctca agcatctggc ggggataaat ccgctctcct ttcaaatcgt tccattcttt      120
gaggcgctgt acagttacgc ccattttttc ggcgatatga tgaagcgtat cccctttccg      180
cactacatat gtaccggtct tcgattcatc gtcatgaagg cggagtgttt ggccggcctt      240
gagatttgaa tgtttcaacc cgtttattct catgatctcc tcgatggata taccgctatc      300
cttgctgatt ctccagagcg tgtccccttt ttgaacggtc accgcaccgc tcattgtccc      360
ggcgttttga taaacgtgga tagaattttg ccggaacgcc tcctcacgaa gcaccgtcag      420
cggattgatt gcatatcttt tatcttcagt ccatgaaccg tgatgcattt caaaatgcag      480
gtgggttccg gtcgatattc                                                  500

<210>  41
<211>  46
<212>  DNA
<213>  Artificial sequence

<220>
<223>  serA1 homology arm 2 forward primer

<400>  41
atgacgctta tttaggagga aatcttacac agaagctgcg gaacct                      46

<210>  42
<211>  41
<212>  DNA
<213>  Artificial sequence

<220>
<223>  serA1 homology arm 2 reverse primer

<400>  42
cagaagaaaa tggaggaatt cgaatatcga ccggaaccca c                           41

<210>  43
<211>  415
<212>  DNA
<213>  Artificial sequence

<220>
<223>  DNA encoding ts1 gRNA expression cassette

<400>  43
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca      240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaat           415

<210>  44
<211>  1000
<212>  DNA
<213>  Artificial sequence

<220>
<223>  serA1 deletion editing template

<400>  44
aatggttctt tcccctgtcc taaacaaaaa acccgcttta ttgaaaaagc ggggctgttt       60
tacagacagg tcaaataaac gtttgaaaat gttcatttca aaacgcgcgg aacctccatc      120
ttctcccatc cagactatac tgtcggcttc ggaatcgcac cgaatcctgc ccataaaaag      180
gctcgcgggc ttagagcgct tgctcatcac cgccggtagg gaatttcacc ctgccccgaa      240
gattgatctt atttattttt aatactgata ttattataaa ttaattgtga aaaaatgtac      300
aggtgcaaag cttattgcgc tgttttggga catcctgcac gatatttcgg taaactcact      360
ttttccgcat actaaaaacc gcacattcac agttatttca tttttaattt tcgtctttcc      420
gcgtgaaact cattgacact ctttatggaa tatggtaaat tatcagatat ttatgacgct      480
tatttaggag gaaatcttac acagaagctg cggaacctga aaagaattcc tttcaggttc      540
cgtttttttt aggaattctc cctgatctca agcatctggc ggggataaat ccgctctcct      600
ttcaaatcgt tccattcttt gaggcgctgt acagttacgc ccattttttc ggcgatatga      660
tgaagcgtat cccctttccg cactacatat gtaccggtct tcgattcatc gtcatgaagg      720
cggagtgttt ggccggcctt gagatttgaa tgtttcaacc cgtttattct catgatctcc      780
tcgatggata taccgctatc cttgctgatt ctccagagcg tgtccccttt ttgaacggtc      840
accgcaccgc tcattgtccc ggcgttttga taaacgtgga tagaattttg ccggaacgcc      900
tcctcacgaa gcaccgtcag cggattgatt gcatatcttt tatcttcagt ccatgaaccg      960
tgatgcattt caaaatgcag gtgggttccg gtcgatattc                           1000

<210>  45
<211>  402
<212>  DNA
<213>  Bacillus  licheniformis

<400>  45
atgacgaact ttggacacca tttacgacaa ttaagggaac ggaaaaaact gaccgtcaat       60
caactggcga tgtattccgg cgtcagttcg gcaggcattt cgcgaatcga aaacggaaag      120
cgcggcgtgc cgaagccggc gacgatcaga aaactggcgg acgctttgaa agtcccgtat      180
gaggaactga tggcatctgc aggctatatc agcgcgtcta cagtccagga agcaagaagc      240
agctatgatt ccatttacga catcgtgtca cagtacgatt tagaggacct ttctctgttt      300
gacagcgaaa agtggaaggt gctttcaaaa aaagacatcg aaaacctgga caaatatttc      360
gactttctcg tgcaggaagc aagcagccga aacaaaaact ga                         402

<210>  46
<211>  20
<212>  DNA
<213>  Bacillus  licheniformis

<400>  46
gatgccatca gttcctcata                                                   20

<210>  47
<211>  23
<212>  DNA
<213>  Bacillus  licheniformis

<400>  47
gatgccatca gttcctcata cgg                                               23

<210>  48
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized DNA encoding variable targeting domain 2

<400>  48
gatgccatca gttcctcata                                                   20

<210>  49
<211>  96
<212>  RNA
<213>  Artificial sequence

<220>
<223>  synthesized gRNA targeting target site 2

<400>  49
gaugccauca guuccucaua guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60
cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96

<210>  50
<211>  500
<212>  DNA
<213>  Bacillus licheniformis

<400>  50
ttgatattca gcaccctgcg catttcgacc gggagaacga ctctgccgag ctcatcgatt       60
ctccggacaa tcccggtatt tttcacgttt gaaaagcctc cttttctcct ttctttattg      120
acttttgtca acatctttat aataaaagag atcttcaaat tttttgttga aatactgaat      180
catctttccg atcacaagtt gtccgggcct cctttcgcca tttaaaactc tgctgagtgt      240
cgccggggat acgccgattt caatggcaag ctgatttaag gagagattgt gttcaatcat      300
gtactggaga acaaaatctc ttttgatatg aatctttttt accatgatta ctcccctttc      360
taatctctta tgtttctttt tatctacatt gaacatatac gatttgttaa cttttgtcaa      420
tacttttacc atccatatgt ttcctatagg caatattcgt actaaaatat tttataataa      480
gagattgcga ggttttggcc                                                  500

<210>  51
<211>  40
<212>  DNA
<213>  artificial sequence

<220>
<223>  rghR1 homology arm 1 forward primer

<400>  51
tgagtaaact tggtctgaca ttgatattca gcaccctgcg                             40

<210>  52
<211>  38
<212>  DNA
<213>  Artificial sequence

<220>
<223>  rghR1 homology arm 1 reverse primer

<400>  52
tgtgccgcgg agaagtatgg ccaaaacctc gcaatctc                               38

<210>  53
<211>  500
<212>  DNA
<213>  Bacillus  licheniformis

<400>  53
atacttctcc gcggcacact ctcctctcta tcattttcgt ctgtttacga tcctgctgtt       60
attttatccc ttatgttaac ttttgtcaat atttttcctg tctaagtatt tcctatagtc      120
aacatttgta ttaaaatgtt catatcatga atttgcgggg gggatggcga tgacaaggtt      180
cggcgagcgg ctcaaagagc tgagggaaca aagaagcctg tcggttaatc agcttgccat      240
gtatgccggt gtgagcgccg cagccatttc cagagccgca gccatttcca gaatcgaaaa      300
cggccaccgc ggcgttccca agcccgcgac gatcagaaaa ttggccgagg ctctgaaaat      360
gccgtacgag cagctcatgg atattgccgg ttatatgaga gctgacgaga ttcgcgaaca      420
gccgcgcggc tatgtcacga tgcaggagat cgcggccaag cacggcgtcg aagacctgtg      480
gctgtttaaa cccgagaaat                                                  500

<210>  54
<211>  38
<212>  DNA
<213>  Artificial sequence

<220>
<223>  rghR1 homology arm 2 forward primer

<400>  54
gagattgcga ggttttggcc atacttctcc gcggcaca                               38

<210>  55
<211>  44
<212>  DNA
<213>  Artificial sequence

<220>
<223>  rghR1 homology arm 2 reverse primer

<400>  55
cagaagaaaa tggaggaatt catttctcgg gtttaaacag ccac                        44

<210>  56
<211>  415
<212>  DNA
<213>  Artificial sequence

<220>
<223>  DNA encoding ts2 expression cassette

<400>  56
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagatgcc atcagttcct      240
catagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaat           415

<210>  57
<211>  1000
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized rghR1 deletion editing template

<400>  57
ttgatattca gcaccctgcg catttcgacc gggagaacga ctctgccgag ctcatcgatt       60
ctccggacaa tcccggtatt tttcacgttt gaaaagcctc cttttctcct ttctttattg      120
acttttgtca acatctttat aataaaagag atcttcaaat tttttgttga aatactgaat      180
catctttccg atcacaagtt gtccgggcct cctttcgcca tttaaaactc tgctgagtgt      240
cgccggggat acgccgattt caatggcaag ctgatttaag gagagattgt gttcaatcat      300
gtactggaga acaaaatctc ttttgatatg aatctttttt accatgatta ctcccctttc      360
taatctctta tgtttctttt tatctacatt gaacatatac gatttgttaa cttttgtcaa      420
tacttttacc atccatatgt ttcctatagg caatattcgt actaaaatat tttataataa      480
gagattgcga ggttttggcc atacttctcc gcggcacact ctcctctcta tcattttcgt      540
ctgtttacga tcctgctgtt attttatccc ttatgttaac ttttgtcaat atttttcctg      600
tctaagtatt tcctatagtc aacatttgta ttaaaatgtt catatcatga atttgcgggg      660
gggatggcga tgacaaggtt cggcgagcgg ctcaaagagc tgagggaaca aagaagcctg      720
tcggttaatc agcttgccat gtatgccggt gtgagcgccg cagccatttc cagagccgca      780
gccatttcca gaatcgaaaa cggccaccgc ggcgttccca agcccgcgac gatcagaaaa      840
ttggccgagg ctctgaaaat gccgtacgag cagctcatgg atattgccgg ttatatgaga      900
gctgacgaga ttcgcgaaca gccgcgcggc tatgtcacga tgcaggagat cgcggccaag      960
cacggcgtcg aagacctgtg gctgtttaaa cccgagaaat                           1000

<210>  58
<211>  1368
<212>  PRT
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155H variant

<400>  58
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile His Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             
Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             
Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             
Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             
Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             
Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             
Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             
Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             
Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             
Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             
Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             
Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             
Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             
Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             
Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             
Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             
Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             
Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             
His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             
Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             
Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             
Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             
Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             
Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             
Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             

<210>  59
<211>  33
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Y155H forward primer

<400>  59
gatctgcgtt taatccatct tgcgttagcg cac                                    33

<210>  60
<211>  33
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Y155H reverse primer

<400>  60
gtgcgctaac gcaagatgga ttaaacgcag atc                                    33

<210>  61
<211>  9724
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF827

<400>  61
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca      240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga      420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag      480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag      540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa      600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta      660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg      720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt      780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac      840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca      900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct      960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt     1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt     1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa     1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat     1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg     1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt     1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca     1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa     1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg     1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc     1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt     1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg     1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg     1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata     1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg     1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga     1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg     1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt     2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc     2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat     2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg     2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg     2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa     2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa     2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta     2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga     2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt     2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc     2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt     2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct     2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt     2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc     2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat     2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag     3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga     3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg     3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt     3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac     3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc     3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt     3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc     3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc     3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg     3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag     3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg     3660
tctgacaaat ggttctttcc cctgtcctaa acaaaaaacc cgctttattg aaaaagcggg     3720
gctgttttac agacaggtca aataaacgtt tgaaaatgtt catttcaaaa cgcgcggaac     3780
ctccatcttc tcccatccag actatactgt cggcttcgga atcgcaccga atcctgccca     3840
taaaaaggct cgcgggctta gagcgcttgc tcatcaccgc cggtagggaa tttcaccctg     3900
ccccgaagat tgatcttatt tatttttaat actgatatta ttataaatta attgtgaaaa     3960
aatgtacagg tgcaaagctt attgcgctgt tttgggacat cctgcacgat atttcggtaa     4020
actcactttt tccgcatact aaaaaccgca cattcacagt tatttcattt ttaattttcg     4080
tctttccgcg tgaaactcat tgacactctt tatggaatat ggtaaattat cagatattta     4140
tgacgcttat ttaggaggaa atcttacaca gaagctgcgg aacctgaaaa gaattccttt     4200
caggttccgt tttttttagg aattctccct gatctcaagc atctggcggg gataaatccg     4260
ctctcctttc aaatcgttcc attctttgag gcgctgtaca gttacgccca ttttttcggc     4320
gatatgatga agcgtatccc ctttccgcac tacatatgta ccggtcttcg attcatcgtc     4380
atgaaggcgg agtgtttggc cggccttgag atttgaatgt ttcaacccgt ttattctcat     4440
gatctcctcg atggatatac cgctatcctt gctgattctc cagagcgtgt cccctttttg     4500
aacggtcacc gcaccgctca ttgtcccggc gttttgataa acgtggatag aattttgccg     4560
gaacgcctcc tcacgaagca ccgtcagcgg attgattgca tatcttttat cttcagtcca     4620
tgaaccgtga tgcatttcaa aatgcaggtg ggttccggtc gatattcgaa ttcctccatt     4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc     4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc     4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc     4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc     4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt     4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa     5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg     5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata     5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac     5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg     5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa     5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta     5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt     5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca     5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg     5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac     5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc     5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt     5760
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg     5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata     5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa     5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa     6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca     6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg     6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag     6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca     6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga     6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg     6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct     6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac     6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc     6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct     6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg     6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca     6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag     6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc     6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag     6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct     6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg     7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga     7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg     7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg     7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc     7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca     7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt     7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg     7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga     7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc     7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag     7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat     7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc     7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac     7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag     7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg     7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca     7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata     8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc     8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag     8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt     8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg     8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt     8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta     8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg     8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg     8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta     8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga     8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag     8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga     8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt     8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag     8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa     8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg     9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg     9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa     9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag     9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt     9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc     9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta     9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac     9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca     9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat     9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac     9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc     9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt     9720
aatc                                                                  9724

<210>  62
<211>  5042
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized Y155H variant expression cassette

<400>  62
attcctccat tttcttctgc tatcaaaata acagactcgt gattttccaa acgagctttc       60
aaaaaagcct ctgccccttg caaatcggat gcctgtctat aaaattcccg atattggtta      120
aacagcggcg caatggcggc cgcatctgat gtctttgctt ggcgaatgtt catcttattt      180
cttcctccct ctcaataatt ttttcattct atcccttttc tgtaaagttt atttttcaga      240
atacttttat catcatgctt tgaaaaaata tcacgataat atccattgtt ctcacggaag      300
cacacgcagg tcatttgaac gaattttttc gacaggaatt tgccgggact caggagcatt      360
taacctaaaa aagcatgaca tttcagcata atgaacattt actcatgtct attttcgttc      420
ttttctgtat gaaaatagtt atttcgagtc tctacggaaa tagcgagaga tgatatacct      480
aaatagagat aaaatcatct caaaaaaatg ggtctactaa aatattattc catctattac      540
aataaattca cagaatagtc ttttaagtaa gtctactctg aattttttta aaaggagagg      600
gtaactagtg gccccaaaaa agaaacgcaa ggttatggat aaaaaataca gcattggtct      660
ggatatcgga accaacagcg ttgggtgggc agtaataaca gatgaataca aagtgccgtc      720
aaaaaaattt aaggttctgg ggaatacaga tcgccacagc ataaaaaaga atctgattgg      780
ggcattgctg tttgattcgg gtgagacagc tgaggccacg cgtctgaaac gtacagcaag      840
aagacgttac acacgtcgta aaaatcgtat ttgctactta caggaaattt tttctaacga      900
aatggccaag gtagatgata gtttcttcca tcgtctcgaa gaatcttttc tggttgagga      960
agataaaaaa cacgaacgtc accctatctt tggcaatatc gtggatgaag tggcctatca     1020
tgaaaaatac cctacgattt atcatcttcg caagaagttg gttgatagta cggacaaagc     1080
ggatctgcgt ttaatccatc ttgcgttagc gcacatgatc aaatttcgtg gtcatttctt     1140
aattgaaggt gatctgaatc ctgataactc tgatgtggac aaattgttta tacaattagt     1200
gcaaacctat aatcagctgt tcgaggaaaa ccccattaat gcctctggag ttgatgccaa     1260
agcgatttta agcgcgagac tttctaagtc ccggcgtctg gagaatctga tcgcccagtt     1320
accaggggaa aagaaaaatg gtctgtttgg taatctgatt gccctcagtc tggggcttac     1380
cccgaacttc aaatccaatt ttgacctggc tgaggacgca aagctgcagc tgagcaaaga     1440
tacttatgat gatgacctcg acaatctgct cgcccagatt ggtgaccaat atgcggatct     1500
gtttctggca gcgaagaatc tttcggatgc tatcttgctg tcggatattc tgcgtgttaa     1560
taccgaaatc accaaagcgc ctctgtctgc aagtatgatc aagagatacg acgagcacca     1620
ccaggacctg actcttctta aggcactggt acgccaacag cttccggaga aatacaaaga     1680
aatattcttc gaccagtcca agaatggtta cgcgggctac atcgatggtg gtgcatcaca     1740
ggaagagttc tataaattta ttaaaccaat ccttgagaaa atggatggca cggaagagtt     1800
acttgttaaa cttaaccgcg aagacttgct tagaaagcaa cgtacattcg acaacggctc     1860
catcccacac cagattcatt taggtgaact tcacgccatc ttgcgcagac aagaagattt     1920
ctatcccttc ttaaaagaca atcgggagaa aatcgagaag atcctgacgt tccgcattcc     1980
ctattatgtc ggtcccctgg cacgtggtaa ttctcggttt gcctggatga cgcgcaaaag     2040
tgaggaaacc atcacccctt ggaactttga agaagtcgtg gataaaggtg ctagcgcgca     2100
gtcttttata gaaagaatga cgaacttcga taaaaacttg cccaacgaaa aagtcctgcc     2160
caagcactct cttttatatg agtactttac tgtgtacaac gaactgacta aagtgaaata     2220
cgttacggaa ggtatgcgca aacctgcctt tcttagtggc gagcagaaaa aagcaattgt     2280
cgatcttctc tttaaaacga atcgcaaggt aactgtaaaa cagctgaagg aagattattt     2340
caaaaagatc gaatgctttg attctgtcga gatctcgggt gtcgaagatc gtttcaacgc     2400
ttccttaggg acctatcatg atttgctgaa gataataaaa gacaaagact ttctcgacaa     2460
tgaagaaaat gaagatattc tggaggatat tgttttgacc ttgaccttat tcgaagatag     2520
agagatgatc gaggagcgct taaaaaccta tgcccacctg tttgatgaca aagtcatgaa     2580
gcaattaaag cgccgcagat atacggggtg gggccgcttg agccgcaagt tgattaacgg     2640
tattagagac aagcagagcg gaaaaactat cctggatttc ctcaaatctg acggatttgc     2700
gaaccgcaat tttatgcagc ttatacatga tgattcgctt acattcaaag aggatattca     2760
gaaggctcag gtgtctgggc aaggtgattc actccacgaa catatagcaa atttggccgg     2820
ctctcctgcg attaagaagg ggatcctgca aacagttaaa gttgtggatg aacttgtaaa     2880
agtaatgggc cgccacaagc cggagaatat cgtgatagaa atggcgcgcg agaatcaaac     2940
gacacaaaaa ggtcaaaaga actcaagaga gagaatgaag cgcattgagg aggggataaa     3000
ggaacttgga tctcaaattc tgaaagaaca tccagttgaa aacactcagc tgcaaaatga     3060
aaaattgtac ctgtactacc tgcagaatgg aagagacatg tacgtggatc aggaattgga     3120
tatcaataga ctctcggact atgacgtaga tcacattgtc cctcagagct tcctcaagga     3180
tgattctata gataataaag tacttacgag atcggacaaa aatcgcggta aatcggataa     3240
cgtcccatcg gaggaagtcg ttaaaaagat gaaaaactat tggcgtcaac tgctgaacgc     3300
caagctgatc acacagcgta agtttgataa tctgactaaa gccgaacgcg gtggtcttag     3360
tgaactcgat aaagcaggat ttataaaacg gcagttagta gaaacgcgcc aaattacgaa     3420
acacgtggct cagatcctcg attctagaat gaatacaaag tacgatgaaa acgataaact     3480
gatccgtgaa gtaaaagtca ttaccttaaa atctaaactt gtgtccgatt tccgcaaaga     3540
ttttcagttt tacaaggtcc gggaaatcaa taactatcac catgcacatg atgcatattt     3600
aaatgcggtt gtaggcacgg cccttattaa gaaataccct aaactcgaaa gtgagtttgt     3660
ttatggggat tataaagtgt atgacgttcg caaaatgatc gcgaaatcag aacaggaaat     3720
cggtaaggct accgctaaat acttttttta ttccaacatt atgaattttt ttaagaccga     3780
aataactctc gcgaatggtg aaatccgtaa acggcctctt atagaaacca atggtgaaac     3840
gggagaaatc gtttgggata aaggtcgtga ctttgccacc gttcgtaaag tcctctcaat     3900
gccgcaagtt aacattgtca agaagacgga agttcaaaca gggggattct ccaaagaatc     3960
tatcctgccg aagcgtaaca gtgataaact tattgccaga aaaaaagatt gggatccaaa     4020
aaaatacgga ggctttgatt cccctaccgt cgcgtatagt gtgctggtgg ttgctaaagt     4080
cgagaaaggg aaaagcaaga aattgaaatc agttaaagaa ctgctgggta ttacaattat     4140
ggaaagatcg tcctttgaga aaaatccgat cgacttttta gaggccaagg ggtataagga     4200
agtgaaaaaa gatctcatca tcaaattacc gaagtatagt ctttttgagc tggaaaacgg     4260
cagaaaaaga atgctggcct ccgcgggcga gttacagaag ggaaatgagc tggcgctgcc     4320
ttccaaatat gttaattttc tgtaccttgc cagtcattat gagaaactga agggcagccc     4380
cgaagataac gaacagaaac aattattcgt ggaacagcat aagcactatt tagatgaaat     4440
tatagagcaa attagtgaat tttctaagcg cgttatcctc gcggatgcta atttagacaa     4500
agtactgtca gcttataata aacatcggga taagccgatt agagaacagg ccgaaaatat     4560
cattcatttg tttaccttaa ccaaccttgg agcaccagct gccttcaaat atttcgatac     4620
cacaattgat cgtaaacggt atacaagtac aaaagaagtc ttggacgcaa ccctcattca     4680
tcaatctatt actggattat atgagacacg cattgatctt tcacagctgg gcggagacaa     4740
gaagaaaaaa ctgaaactgc accatcatca ccatcatcat caccatcatt gataaacata     4800
aaaaaccggc cttggccccg ccggtttttt attatttttc ttcctccgca tgttcaatcc     4860
gctccataat cgacggatgg ctccctctga aaattttaac gagaaacggc gggttgaccc     4920
ggctcagtcc cgtaacggcc aagtcctgaa acgtctcaat cgccgcttcc cggtttccgg     4980
tcagctcaat gccgtaacgg tcggcggcgt tttcctgata ccgggagacg gcattcgtaa     5040
tc                                                                    5042

<210>  63
<211>  9724
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF856

<400>  63
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagatgcc atcagttcct      240
catagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga      420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag      480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag      540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa      600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta      660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg      720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt      780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac      840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca      900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct      960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt     1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt     1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa     1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat     1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg     1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt     1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca     1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa     1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg     1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc     1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt     1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg     1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg     1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata     1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg     1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga     1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg     1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt     2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc     2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat     2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg     2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg     2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa     2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa     2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta     2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga     2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt     2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc     2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt     2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct     2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt     2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc     2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat     2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag     3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga     3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg     3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt     3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac     3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc     3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt     3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc     3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc     3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg     3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag     3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg     3660
tctgacattg atattcagca ccctgcgcat ttcgaccggg agaacgactc tgccgagctc     3720
atcgattctc cggacaatcc cggtattttt cacgtttgaa aagcctcctt ttctcctttc     3780
tttattgact tttgtcaaca tctttataat aaaagagatc ttcaaatttt ttgttgaaat     3840
actgaatcat ctttccgatc acaagttgtc cgggcctcct ttcgccattt aaaactctgc     3900
tgagtgtcgc cggggatacg ccgatttcaa tggcaagctg atttaaggag agattgtgtt     3960
caatcatgta ctggagaaca aaatctcttt tgatatgaat cttttttacc atgattactc     4020
ccctttctaa tctcttatgt ttctttttat ctacattgaa catatacgat ttgttaactt     4080
ttgtcaatac ttttaccatc catatgtttc ctataggcaa tattcgtact aaaatatttt     4140
ataataagag attgcgaggt tttggccata cttctccgcg gcacactctc ctctctatca     4200
ttttcgtctg tttacgatcc tgctgttatt ttatccctta tgttaacttt tgtcaatatt     4260
tttcctgtct aagtatttcc tatagtcaac atttgtatta aaatgttcat atcatgaatt     4320
tgcggggggg atggcgatga caaggttcgg cgagcggctc aaagagctga gggaacaaag     4380
aagcctgtcg gttaatcagc ttgccatgta tgccggtgtg agcgccgcag ccatttccag     4440
agccgcagcc atttccagaa tcgaaaacgg ccaccgcggc gttcccaagc ccgcgacgat     4500
cagaaaattg gccgaggctc tgaaaatgcc gtacgagcag ctcatggata ttgccggtta     4560
tatgagagct gacgagattc gcgaacagcc gcgcggctat gtcacgatgc aggagatcgc     4620
ggccaagcac ggcgtcgaag acctgtggct gtttaaaccc gagaaatgaa ttcctccatt     4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc     4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc     4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc     4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc     4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt     4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa     5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg     5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata     5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac     5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg     5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa     5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta     5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt     5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca     5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg     5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac     5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc     5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt     5760
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg     5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata     5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa     5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa     6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca     6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg     6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag     6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca     6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga     6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg     6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct     6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac     6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc     6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct     6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg     6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca     6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag     6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc     6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag     6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct     6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg     7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga     7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg     7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg     7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc     7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca     7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt     7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg     7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga     7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc     7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag     7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat     7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc     7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac     7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag     7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg     7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca     7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata     8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc     8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag     8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt     8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg     8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt     8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta     8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg     8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg     8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta     8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga     8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag     8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga     8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt     8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag     8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa     8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg     9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg     9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa     9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag     9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt     9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc     9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta     9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac     9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca     9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat     9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac     9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc     9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt     9720
aatc                                                                  9724

<210>  64
<211>  6393
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized pBL.comK-syn

<400>  64
aagcttcata tgcaagggtt tattgttttc taaaatctga ttaccaatta gaatgaatat       60
ttcccaaata ttaaataata aaacaaaaaa attgaaaaaa gtgtttccac cattttttca      120
atttttttat aattttttta atctgttatt taaatagttt atagttaaat ttacattttc      180
attagtccat tcaatattct ctccaagata actacgaact gctaacaaaa ttctctccct      240
atgttctaat ggagaagatt cagccactgc atttcccgca atatcttttg gtatgatttt      300
acccgtgtcc atagttaaaa tcatacggca taaagttaat atagagttgg tttcatcatc      360
ctgataatta tctattaatt cctctgacga atccataatg gctcttctca catcagaaaa      420
tggaatatca ggtagtaatt cctctaagtc ataatttccg tatattcttt tattttttcg      480
ttttgcttgg taaagcatta tggttaaatc tgaatttaat tccttctgag gaatgtatcc      540
ttgttcataa agctcttgta accattctcc ataaataaat tcttgtttgg gaggatgatt      600
ccacggtacc atttcttgct gaataataat tgttaattca atatatcgta agttgctttt      660
atctcctatt ttttttgaaa taggtctaat tttttgtata agtatttctt tactttgatc      720
tgtcaatggt tcagatacga cgactaaaaa gtcaagatca ctatttggtt ttagtccact      780
ctcaactcct gatccaaaca tgtaagtacc aataaggtta ttttttaaat gtttccgaag      840
tatttttttc actttattaa tttgttcgta tgtattcaaa tatatcctcc tcactatttt      900
gattagtacc tattttatat ccatagttgt taattaaata aacttaattt agtttattta      960
tggatttcat tggcttctaa attttttatc tagataataa ttattttagt taattttatt     1020
ctagattata tatgatatga tctttcattt ccataaaact aaagtaagtg taaacctatt     1080
cattgtttta aaaatatctc ttgccagtca cgttacgtta ttagttatag ttattataac     1140
atgtattcac gaacgggcgc gccggtatcc gcgcttcttg agcactattt attcaaagcc     1200
gctccagatc aatagcgctt tttcagctcc ctgaggatga attcgtatat cagctgattc     1260
cggtcttctt tcggatagag cataaattcc tgtttcttct gcatggggtt tccttcaatc     1320
ctgtcgataa attttgttct cagccatgcc gttcggtaaa cctggttttc gaaagatgag     1380
atggatacgg gcagctccag cgtttccccg ttgacaaacg tgacaaacgt gttgtcatac     1440
tttgccgcgc aaaactcgtg aacatgcgca tgggaaagcc acccgcactg aggacgagtt     1500
gaggaaaatg tggggaaaag aaaaatgttg tttgagtgat ccaccatgat cggcggttta     1560
tgggaaactt taatgacttc atatgtgccc gcttttcttc ccgcatagct cgatccgaaa     1620
tagcggcagc ttctttcgat aatttgaaac ggcttcatat tgacgcggaa agtcctgtcg     1680
gtctcaagta tttttgaggc ggatttctcc ccctcaccca gaggcaggac agccattgtc     1740
gaactgttta cttcatacgt atcctttgtc atatcctctg tgctcatgtg atttccccct     1800
taaaaataaa ttcattcaaa tacagatgca ttttatttca tatagtaagt acatcaccta     1860
ttagtttgtt gtttaaacaa actaacttat tttcatctta tataacctcg tcagtatttt     1920
caatattttt tttagttttt tatgaacaca ttagatttaa taaagggaag attcgctatg     1980
tactatgttg atacttaatt taaagattaa acaaatggag tggatgaagt ggatatcgct     2040
gatcaaacct ttgtcaaaaa agtaaatcaa aagttattat taaaagaaat ccttaaaaat     2100
tcacctattt caagagcaaa attatctgaa atgactggat taaataaatc aactgtctca     2160
tcacaggtaa acacgttaat gaaagaaagt atggtatttg aaataggtca aggacaatca     2220
agtggcggaa gaagacctgt catgcttgtt tttaataaaa aggcaggata ctccgttgga     2280
atagatgttg gtgtggatta tattaatggc attttaacag accttgaagg aacaatcgtt     2340
cttgatcaat accgccattt ggaatccaat tctccagaaa taacgaaaga cattttgatt     2400
gatatgattc atcactttat tacgcaaatg ccccaatctc cgtacgggtt tattggtata     2460
ggtacttgcg tgcctggact cattgataaa gatcaaaaaa ttgttttcac tccgaactcc     2520
aactggagag atattgactt aaaatcttcg atacaagaga agtacaatgt gtctgttttt     2580
attgaaaatg aggcaaatgc tggcgcatat ggagaaaaac tatttggagc tgcaaaaaat     2640
cacgataaca ttatttacgt aagtatcagc acaggaatag ggatcggtgt tattatcaac     2700
aatcatttat atagaggagt aagcggcttc tctggagaaa tgggacatat gacaatagac     2760
tttaatggtc ctaaatgcag ttgcggaaac cgaggatgct gggaattgta tgcttcagag     2820
aaggctttat ttaaatctct tcagaccaaa gagaaaaaac tgtcctatca agatatcata     2880
aacctcgccc atctgaatga tatcggaacc ttaaatgcat tacaaaattt tggattctat     2940
ttaggaatag gccttaccaa tattctaaat actctcaacc cacaagccgt aattttaaga     3000
aatagcataa ttgaatcgca tcctatggtt ttaaattcaa tgagaagtga agtatcatca     3060
agggtttatt cccaattagg caatagctat gaattattgc catcttcctt aggacagaat     3120
gcaccggcat taggaatgtc ctccattgtg attgatcatt ttctggacat gattacaatg     3180
taatttttta tggaatggac agctcatctt taaagatgag tttttttatt ctaggagtat     3240
ttctgaagca atagtgacat ggcaccttct catatgaaaa aggagttcta aaataaaaat     3300
ctcctttttc atgtgcaaat tatttttctt tataacgaaa atatctaaat gacaatgcat     3360
atgcaagagg ggatcacata aatatatatt ttaaaaatat cccactttat ccaattttcg     3420
tttgttgaac taatgggtgc tttagttgaa gaataaaaga ccacattaaa aaatgtggtc     3480
ttttgtgttt ttttaaagga tttgagcgta gcgaaaaatc cttttctttc ttatcttgat     3540
actatataga aacaacatca tttttcaaaa ttaggtcaaa gccttgtgta tcaagggttt     3600
gatggttctt tgacaggtaa aaactccttc tgctattatt aaatactata tagaaacaac     3660
atcatttttc aaaattaggt caaagccttg tgtatcaagg gtttgatggt tctttgacag     3720
gtaaaaactc cttctgctat tattaaggtg tcgaatcaaa ataatagaat gctagagaac     3780
tagctcagaa ggagtttttt tgttgattta ttcatctgaa aatgattata gcatcctcga     3840
agataaaacc gcaacaggta aaaagcggga ttggaagggg aaaaagagac ggacgaacct     3900
catggcggag cattacgaag cgttagagag taagattggg gcaccttact atggcaaaaa     3960
ggctgaaaaa ctaattagtt gtgcagagta tctttcgttt aagagagacc cggagacggg     4020
caagttaaaa ctgtatcaag cccatttttg taaagtgagg ttatgtccga tgtgtgcgtg     4080
gcgcaggtcg ttaaaaattg cttatcacaa taagttgatc gtagaggaag ccaatagaca     4140
gtacggctgc ggatggattt ttctcacgct gacgattcga aatgtaaagg gagaacggct     4200
gaagccacaa atttctgcga tgatggaagg ctttaggaaa ctgttccagt acaaaaaagt     4260
aaaaacttcg gttcttggat ttttcagagc tttagagatt accaaaaatc atgaagaaga     4320
tacatatcat cctcattttc atgtgttgat accagtaagg aaaaattatt ttgggaaaaa     4380
ctatattaag caggcggagt ggacgagcct ttggaaaaag gcgatgaaat tggattacac     4440
tccaattgtc gatattcgtc gagtgaaagg taaagctaag attgacgctg aacagattga     4500
aaacgatgtg cggaacgcaa tgatggagca aaaagctgtt ctcgaaatct ctaaatatcc     4560
ggttaaggat acggatgttg tgcgcggtaa taaggtgact gaagacaatc tgaacacggt     4620
gctttacttg gatgatgcgt tggcagctcg aaggttaatt ggatacggtg gcattttgaa     4680
ggagatacat aaagagctga atcttggtga tgcggaggac ggcgatctgg tcaagattga     4740
ggaagaagat gacgaggttg caaatggtgc atttgaggtt atggcttatt ggcatcctgg     4800
cattaaaaat tacataatca aataaaaaaa gcagaccttt agaaggcctg cttttttaac     4860
taacccattt gtattgtgtt gaaatatgtt ttgtatggtg cactctcagt acaatctgct     4920
ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac     4980
gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca     5040
tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac     5100
gcctattttt ataggttaat gtcatgataa taatggtttc ttagcgattc acaaaaaata     5160
ggcacacgaa aaacaagtta agggatgcag tttatgcatc ccttaactta aaatactaaa     5220
aatgcccata ttttttcctc cttataaaat tagtataatt atagcacgag atctaaaagg     5280
atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg     5340
ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt     5400
ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg     5460
ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata     5520
ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca     5580
ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag     5640
tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc     5700
tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga     5760
tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg     5820
tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac     5880
gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg     5940
tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg     6000
ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct     6060
gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc     6120
gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc     6180
cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg     6240
ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta     6300
cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca     6360
ggaaacagct atgaccatga ttacgccgga tcc                                  6393

<210>  65
<211>  2800
<212>  DNA
<213>  Bacillus licheniformis

<400>  65
tagagacgag acgtctcacc ttgttgtgtt tcattttgtc actctctcct tttcgatcac       60
atctcacgaa aagaggaatg gttctttccc ctgtcctaaa caaaaaaccc gctttattga      120
aaaagcgggg ctgttttaca gacaggtcaa ataaacgttt gaaaatgttc atttcaaaac      180
gcgcggaacc tccatcttct cccatccaga ctatactgtc ggcttcggaa tcgcaccgaa      240
tcctgcccat aaaaaggctc gcgggcttag agcgcttgct catcaccgcc ggtagggaat      300
ttcaccctgc cccgaagatt gatcttattt atttttaata ctgatattat tataaattaa      360
ttgtgaaaaa atgtacaggt gcaaagctta ttgcgctgtt ttgggacatc ctgcacgata      420
tttcggtaaa ctcacttttt ccgcatacta aaaaccgcac attcacagtt atttcatttt      480
taattttcgt ctttccgcgt gaaactcatt gacactcttt atggaatatg gtaaattatc      540
agatatttat gacgcttatt taggaggaaa tcttacatgt ttcgagtatt ggtctcagat      600
aaaatgtcca gcgacggcct caaaccatta atggaagcag attttattga aattgtagaa      660
aagaatgttg cggaagcgga agacgagctt catacgtttg acgcgctctt ggtgcggagc      720
gccacgaagg taaccgaaga gctgtttaaa aagatgactt cgctgaaaat cgtcgccaga      780
gcaggtgtcg gcgtcgacaa tatcgatatt gacgaggcga caaaacacgg tgttatcgtc      840
gtaaacgcgc caaacgggaa tacaatttca accgctgaac atacctttgc aatgttttca      900
gcgttaatga gacatattcc gcaggcaaac atctccgtga aatcaaggga gtggaatcgt      960
tcggcttacg tcggttcaga gctttacgga aaaacgctcg gcatcatcgg aatgggccgc     1020
atcggaagcg aaatcgcgag ccgcgcaaaa gcattcggta tgaccgttca tgtatttgac     1080
ccgttcctga cccaagaaag ggcaagcaag ctcggcgtta acgcgaacag ctttgaagaa     1140
gttctggcat gcgccgacat cattacggtt cataccccgc tcacgaaaga aacgaaggga     1200
cttttgaaca aagaaaccat cgcaaaaacg aaaaaaggcg ttcgtctcgt taactgtgca     1260
agaggcggca tcatcgatga agcagcgctt ttggaagctc tggaaagcgg acatgtcgct     1320
ggcgctgcct tggatgtatt cgaagtcgag cctccggtcg attcaaaact gatcgatcat     1380
ccgcttgtag tcgcgactcc tcacttgggc gcctcaacaa aagaagccca gctgaatgtc     1440
gctgcacaag tgtccgaaga agtccttcag tatgcgcaag gaaaccctgt gatgtccgcg     1500
atcaaccttc cggccatgac aaaggattca ttcgaaaaaa tccagcctta tcatcagttt     1560
gccaatacga tcggaaacct tgtgtctcag tgcatgaatg agcctgttca agatgtagcc     1620
atccaatatg aaggctccat cgccaaactt gaaacgtcat ttattacgaa aagccttttg     1680
gccggatttc tgaagccgag ggtcgcggct accgttaacg aagtgaatgc cggcaccgtt     1740
gcgaaagagc gcggcatcag cttcagcgaa aaaatttctt ccaatgagtc aggctatgaa     1800
aactgcatct ctgtgactgt cacgggagat gtaacaacat tctctttaag agcgacgtac     1860
attccgcact tcggcggacg catcgttgcc ttaaacggct ttgatattga tttttatccg     1920
gctggacacc ttgtctacat tcaccaccag gataaaccag gggctatcgg ccatgtcgga     1980
cgaattttag gagaccatga catcaatatc gccactatgc aggtaggccg aaaagaaaaa     2040
ggcggagaag cgatcatgat gctttccttt gaccgccacc ttgaggacga tattttagct     2100
gagctgaaaa acatcccgga tatcgtgtct gttaaagcca tcgaccttcc ttaaacagaa     2160
gctgcggaac ctgaaaagaa ttcctttcag gttccgtttt ttttaggaat tctccctgat     2220
ctcaagcatc tggcggggat aaatccgctc tcctttcaaa tcgttccatt ctttgaggcg     2280
ctgtacagtt acgcccattt tttcggcgat atgatgaagc gtatcccctt tccgcactac     2340
atatgtaccg gtcttcgatt catcgtcatg aaggcggagt gtttggccgg ccttgagatt     2400
tgaatgtttc aacccgttta ttctcatgat ctcctcgatg gatataccgc tatccttgct     2460
gattctccag agcgtgtccc ctttttgaac ggtcaccgca ccgctcattg tcccggcgtt     2520
ttgataaacg tggatagaat tttgccggaa cgcctcctca cgaagcaccg tcagcggatt     2580
gattgcatat cttttatctt cagtccatga accgtgatgc atttcaaaat gcaggtgggt     2640
tccggtcgat attcccgtat tgccgatgat tccgatttgc tcgccttttt tcacccgctc     2700
cttttccttt ttcaggcgtt tgcttaagtg ggcataaacg gtttcatatc cgttgtcatg     2760
tttaataaat atcacttggc cgtaggagtc ggattgatac                           2800

<210>  66
<211>  1222
<212>  DNA
<213>  Artificial sequence

<220>
<223>  target site 1 edited locus

<400>  66
tagagacgag acgtctcacc ttgttgtgtt tcattttgtc actctctcct tttcgatcac       60
atctcacgaa aagaggaatg gttctttccc ctgtcctaaa caaaaaaccc gctttattga      120
aaaagcgggg ctgttttaca gacaggtcaa ataaacgttt gaaaatgttc atttcaaaac      180
gcgcggaacc tccatcttct cccatccaga ctatactgtc ggcttcggaa tcgcaccgaa      240
tcctgcccat aaaaaggctc gcgggcttag agcgcttgct catcaccgcc ggtagggaat      300
ttcaccctgc cccgaagatt gatcttattt atttttaata ctgatattat tataaattaa      360
ttgtgaaaaa atgtacaggt gcaaagctta ttgcgctgtt ttgggacatc ctgcacgata      420
tttcggtaaa ctcacttttt ccgcatacta aaaaccgcac attcacagtt atttcatttt      480
taattttcgt ctttccgcgt gaaactcatt gacactcttt atggaatatg gtaaattatc      540
agatatttat gacgcttatt taggaggaaa tcttacacag aagctgcgga acctgaaaag      600
aattcctttc aggttccgtt ttttttagga attctccctg atctcaagca tctggcgggg      660
ataaatccgc tctcctttca aatcgttcca ttctttgagg cgctgtacag ttacgcccat      720
tttttcggcg atatgatgaa gcgtatcccc tttccgcact acatatgtac cggtcttcga      780
ttcatcgtca tgaaggcgga gtgtttggcc ggccttgaga tttgaatgtt tcaacccgtt      840
tattctcatg atctcctcga tggatatacc gctatccttg ctgattctcc agagcgtgtc      900
ccctttttga acggtcaccg caccgctcat tgtcccggcg ttttgataaa cgtggataga      960
attttgccgg aacgcctcct cacgaagcac cgtcagcgga ttgattgcat atcttttatc     1020
ttcagtccat gaaccgtgat gcatttcaaa atgcaggtgg gttccggtcg atattcccgt     1080
attgccgatg attccgattt gctcgccttt tttcacccgc tccttttcct ttttcaggcg     1140
tttgcttaag tgggcataaa cggtttcata tccgttgtca tgtttaataa atatcacttg     1200
gccgtaggag tcggattgat ac                                              1222

<210>  67
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  67
tagagacgag acgtctcacc                                                   20

<210>  68
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  68
gtatcaatcc gactcctacg g                                                 21

<210>  69
<211>  1517
<212>  DNA
<213>  Bacillus licheniformis

<400>  69
atcaaacatg ccatgtttgc ggcgtatttt gtcaaaatga tattttcgcc gtcggtatat       60
atttcgagcg ggtccttttc attgatattc agcaccctgc gcatttcgac cgggagaacg      120
actctgccga gctcatcgat tctccggaca atcccggtat ttttcacgtt tgaaaagcct      180
ccttttctcc tttctttatt gacttttgtc aacatcttta taataaaaga gatcttcaaa      240
ttttttgttg aaatactgaa tcatctttcc gatcacaagt tgtccgggcc tcctttcgcc      300
atttaaaact ctgctgagtg tcgccgggga tacgccgatt tcaatggcaa gctgatttaa      360
ggagagattg tgttcaatca tgtactggag aacaaaatct cttttgatat gaatcttttt      420
taccatgatt actccccttt ctaatctctt atgtttcttt ttatctacat tgaacatata      480
cgatttgtta acttttgtca atacttttac catccatatg tttcctatag gcaatattcg      540
tactaaaata ttttataata agagattgcg aggttttggc catgacgaac tttggacacc      600
atttacgaca attaagggaa cggaaaaaac tgaccgtcaa tcaactggcg atgtattccg      660
gcgtcagttc ggcaggcatt tcgcgaatcg aaaacggaaa gcgcggcgtg ccgaagccgg      720
cgacgatcag aaaactggcg gacgctttga aagtcccgta tgaggaactg atggcatctg      780
caggctatat cagcgcgtct acagtccagg aagcaagaag cagctatgat tccatttacg      840
acatcgtgtc acagtacgat ttagaggacc tttctctgtt tgacagcgaa aagtggaagg      900
tgctttcaaa aaaagacatc gaaaacctgg acaaatattt cgactttctc gtgcaggaag      960
caagcagccg aaacaaaaac tgaatacttc tccgcggcac actctcctct ctatcatttt     1020
cgtctgttta cgatcctgct gttattttat cccttatgtt aacttttgtc aatatttttc     1080
ctgtctaagt atttcctata gtcaacattt gtattaaaat gttcatatca tgaatttgcg     1140
ggggggatgg cgatgacaag gttcggcgag cggctcaaag agctgaggga acaaagaagc     1200
ctgtcggtta atcagcttgc catgtatgcc ggtgtgagcg ccgcagccat ttccagagcc     1260
gcagccattt ccagaatcga aaacggccac cgcggcgttc ccaagcccgc gacgatcaga     1320
aaattggccg aggctctgaa aatgccgtac gagcagctca tggatattgc cggttatatg     1380
agagctgacg agattcgcga acagccgcgc ggctatgtca cgatgcagga gatcgcggcc     1440
aagcacggcg tcgaagacct gtggctgttt aaacccgaga aatgggactg tttgtcccgc     1500
gaagacctgc tcaacct                                                    1517

<210>  70
<211>  1115
<212>  DNA
<213>  Artificial sequence

<220>
<223>  target site 2 edited locus

<400>  70
atcaaacatg ccatgtttgc ggcgtatttt gtcaaaatga tattttcgcc gtcggtatat       60
atttcgagcg ggtccttttc attgatattc agcaccctgc gcatttcgac cgggagaacg      120
actctgccga gctcatcgat tctccggaca atcccggtat ttttcacgtt tgaaaagcct      180
ccttttctcc tttctttatt gacttttgtc aacatcttta taataaaaga gatcttcaaa      240
ttttttgttg aaatactgaa tcatctttcc gatcacaagt tgtccgggcc tcctttcgcc      300
atttaaaact ctgctgagtg tcgccgggga tacgccgatt tcaatggcaa gctgatttaa      360
ggagagattg tgttcaatca tgtactggag aacaaaatct cttttgatat gaatcttttt      420
taccatgatt actccccttt ctaatctctt atgtttcttt ttatctacat tgaacatata      480
cgatttgtta acttttgtca atacttttac catccatatg tttcctatag gcaatattcg      540
tactaaaata ttttataata agagattgcg aggttttggc catacttctc cgcggcacac      600
tctcctctct atcattttcg tctgtttacg atcctgctgt tattttatcc cttatgttaa      660
cttttgtcaa tatttttcct gtctaagtat ttcctatagt caacatttgt attaaaatgt      720
tcatatcatg aatttgcggg ggggatggcg atgacaaggt tcggcgagcg gctcaaagag      780
ctgagggaac aaagaagcct gtcggttaat cagcttgcca tgtatgccgg tgtgagcgcc      840
gcagccattt ccagagccgc agccatttcc agaatcgaaa acggccaccg cggcgttccc      900
aagcccgcga cgatcagaaa attggccgag gctctgaaaa tgccgtacga gcagctcatg      960
gatattgccg gttatatgag agctgacgag attcgcgaac agccgcgcgg ctatgtcacg     1020
atgcaggaga tcgcggccaa gcacggcgtc gaagacctgt ggctgtttaa acccgagaaa     1080
tgggactgtt tgtcccgcga agacctgctc aacct                                1115

<210>  71
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  71
atcaaacatg ccatgtttgc                                                   20

<210>  72
<211>  18
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  72
aggttgagca ggtcttcg                                                     18

<210>  73
<211>  4272
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Yarrowia codon optimized Cas9

<400>  73
atggggggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa       60
atgggtcggg atctgtacga cgatgacgat aaggatccga gctcgagatc tgcagctggt      120
accatatggg aattcgacaa gaaatactcc atcggcctgg acattggaac caactctgtc      180
ggctgggctg tcatcaccga cgagtacaag gtgccctcca agaaattcaa ggtcctcgga      240
aacaccgatc gacactccat caagaaaaac ctcattggtg ccctgttgtt cgattctggc      300
gagactgccg aagctaccag actcaagcga actgctcggc gacgttacac ccgacggaag      360
aaccgaatct gctacctgca ggagatcttt tccaacgaga tggccaaggt ggacgattcg      420
ttctttcatc gactggagga atccttcctc gtcgaggaag acaagaaaca cgagcgtcat      480
cccatctttg gcaacattgt ggacgaggtt gcttaccacg agaagtatcc taccatctac      540
cacctgcgaa agaaactcgt cgattccacc gacaaggcgg atctcagact tatctacctc      600
gctctggcac acatgatcaa gtttcgaggt catttcctca tcgagggcga tctcaatccc      660
gacaacagcg atgtggacaa gctgttcatt cagctcgttc agacctacaa ccagctgttc      720
gaggaaaacc ccatcaatgc ctccggagtc gatgcaaagg ccatcttgtc tgctcgactc      780
tcgaagagca gacgactgga gaacctcatt gcccaacttc ctggcgagaa aaagaacgga      840
ctgtttggca acctcattgc cctttctctt ggtctcacac ccaacttcaa gtccaacttc      900
gatctggcgg aggacgccaa gctccagctg tccaaggaca cctacgacga tgacctcgac      960
aacctgcttg cacagattgg cgatcagtac gccgacctgt ttctcgctgc caagaacctt     1020
tcggatgcta ttctcttgtc tgacattctg cgagtcaaca ccgagatcac aaaggctccc     1080
ctttctgcct ccatgatcaa gcgatacgac gagcaccatc aggatctcac actgctcaag     1140
gctcttgtcc gacagcaact gcccgagaag tacaaggaga tctttttcga tcagtcgaag     1200
aacggctacg ctggatacat cgacggcgga gcctctcagg aagagttcta caagttcatc     1260
aagccaattc tcgagaagat ggacggaacc gaggaactgc ttgtcaagct caatcgagag     1320
gatctgcttc ggaagcaacg aaccttcgac aacggcagca ttcctcatca gatccacctc     1380
ggtgagctgc acgccattct tcgacgtcag gaagacttct acccctttct caaggacaac     1440
cgagagaaga tcgagaagat tcttaccttt cgaatcccct actatgttgg tcctcttgcc     1500
agaggaaact ctcgatttgc ttggatgact cgaaagtccg aggaaaccat cactccctgg     1560
aacttcgagg aagtcgtgga caagggtgcc tctgcacagt ccttcatcga gcgaatgacc     1620
aacttcgaca agaatctgcc caacgagaag gttcttccca agcattcgct gctctacgag     1680
tactttacag tctacaacga actcaccaaa gtcaagtacg ttaccgaggg aatgcgaaag     1740
cctgccttct tgtctggcga acagaagaaa gccattgtcg atctcctgtt caagaccaac     1800
cgaaaggtca ctgttaagca gctcaaggag gactacttca agaaaatcga gtgtttcgac     1860
agcgtcgaga tttccggagt tgaggaccga ttcaacgcct ctttgggcac ctatcacgat     1920
ctgctcaaga ttatcaagga caaggatttt ctcgacaacg aggaaaacga ggacattctg     1980
gaggacatcg tgctcactct taccctgttc gaagatcggg agatgatcga ggaacgactc     2040
aagacatacg ctcacctgtt cgacgacaag gtcatgaaac aactcaagcg acgtagatac     2100
accggctggg gaagactttc gcgaaagctc atcaacggca tcagagacaa gcagtccgga     2160
aagaccattc tggactttct caagtccgat ggctttgcca accgaaactt catgcagctc     2220
attcacgacg attctcttac cttcaaggag gacatccaga aggcacaagt gtccggtcag     2280
ggcgacagct tgcacgaaca tattgccaac ctggctggtt cgccagccat caagaaaggc     2340
attctccaga ctgtcaaggt tgtcgacgag ctggtgaagg tcatgggacg tcacaagccc     2400
gagaacattg tgatcgagat ggccagagag aaccagacaa ctcaaaaggg tcagaaaaac     2460
tcgcgagagc ggatgaagcg aatcgaggaa ggcatcaagg agctgggatc ccagattctc     2520
aaggagcatc ccgtcgagaa cactcaactg cagaacgaga agctgtatct ctactatctg     2580
cagaatggtc gagacatgta cgtggatcag gaactggaca tcaatcgtct cagcgactac     2640
gatgtggacc acattgtccc tcaatccttt ctcaaggacg attctatcga caacaaggtc     2700
cttacacgat ccgacaagaa cagaggcaag tcggacaacg ttcccagcga agaggtggtc     2760
aaaaagatga agaactactg gcgacagctg ctcaacgcca agctcattac ccagcgaaag     2820
ttcgacaatc ttaccaaggc cgagcgaggc ggtctgtccg agctcgacaa ggctggcttc     2880
atcaagcgtc aactcgtcga gaccagacag atcacaaagc acgtcgcaca gattctcgat     2940
tctcggatga acaccaagta cgacgagaac gacaagctca tccgagaggt caaggtgatt     3000
actctcaagt ccaaactggt ctccgatttc cgaaaggact ttcagttcta caaggtgcga     3060
gagatcaaca attaccacca tgcccacgat gcttacctca acgccgtcgt tggcactgcg     3120
ctcatcaaga aataccccaa gctcgaaagc gagttcgttt acggcgatta caaggtctac     3180
gacgttcgaa agatgattgc caagtccgaa caggagattg gcaaggctac tgccaagtac     3240
ttcttttact ccaacatcat gaactttttc aagaccgaga tcaccttggc caacggagag     3300
attcgaaaga gaccacttat cgagaccaac ggcgaaactg gagagatcgt gtgggacaag     3360
ggtcgagact ttgcaaccgt gcgaaaggtt ctgtcgatgc ctcaggtcaa catcgtcaag     3420
aaaaccgagg ttcagactgg cggattctcc aaggagtcga ttctgcccaa gcgaaactcc     3480
gacaagctca tcgctcgaaa gaaagactgg gatcccaaga aatacggtgg cttcgattct     3540
cctaccgtcg cctattccgt gcttgtcgtt gcgaaggtcg agaagggcaa gtccaaaaag     3600
ctcaagtccg tcaaggagct gctcggaatt accatcatgg agcgatcgag cttcgagaag     3660
aatcccatcg acttcttgga agccaagggt tacaaggagg tcaagaaaga cctcattatc     3720
aagctgccca agtactctct gttcgaactg gagaacggtc gaaagcgtat gctcgcctcc     3780
gctggcgagc tgcagaaggg aaacgagctt gccttgcctt cgaagtacgt caactttctc     3840
tatctggctt ctcactacga gaagctcaag ggttctcccg aggacaacga acagaagcaa     3900
ctcttcgttg agcagcacaa acattacctc gacgagatta tcgagcagat ttccgagttt     3960
tcgaagcgag tcatcctggc tgatgccaac ttggacaagg tgctctctgc ctacaacaag     4020
catcgggaca aacccattcg agaacaggcg gagaacatca ttcacctgtt tactcttacc     4080
aacctgggtg ctcctgcagc tttcaagtac ttcgatacca ctatcgaccg aaagcggtac     4140
acatccacca aggaggttct cgatgccacc ctgattcacc agtccatcac tggcctgtac     4200
gagacccgaa tcgacctgtc tcagcttggt ggcgactcca gagccgatcc caagaaaaag     4260
cgaaaggtct aa                                                         4272

<210>  74
<211>  9
<212>  PRT
<213>  Simian virus 40

<400>  74
Met Ala Pro Lys Lys Lys Arg Lys Val 
1               5                   

<210>  75
<211>  546
<212>  DNA
<213>  Yarrowia lipolytica

<400>  75
tcgacgttta aaccatcatc taagggcctc aaaactacct cggaactgct gcgctgatct       60
ggacaccaca gaggttccga gcactttagg ttgcaccaaa tgtcccacca ggtgcaggca      120
gaaaacgctg gaacagcgtg tacagtttgt cttaacaaaa agtgagggcg ctgaggtcga      180
gcagggtggt gtgacttgtt atagccttta gagctgcgaa agcgcgtatg gatttggctc      240
atcaggccag attgagggtc tgtggacaca tgtcatgtta gtgtacttca atcgccccct      300
ggatatagcc ccgacaatag gccgtggcct catttttttg ccttccgcac atttccattg      360
ctcggtaccc acaccttgct tctcctgcac ttgccaacct taatactggt ttacattgac      420
caacatctta caagcggggg gcttgtctag ggtatatata aacagtggct ctcccaatcg      480
gttgccagtc tcttttttcc tttctttccc cacagattcg aaatctaaac tacacatcac      540
accatg                                                                 546

<210>  76
<211>  4683
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Yarrowia Cas9 expression cassette

<400>  76
tcgacgttta aaccatcatc taagggcctc aaaactacct cggaactgct gcgctgatct       60
ggacaccaca gaggttccga gcactttagg ttgcaccaaa tgtcccacca ggtgcaggca      120
gaaaacgctg gaacagcgtg tacagtttgt cttaacaaaa agtgagggcg ctgaggtcga      180
gcagggtggt gtgacttgtt atagccttta gagctgcgaa agcgcgtatg gatttggctc      240
atcaggccag attgagggtc tgtggacaca tgtcatgtta gtgtacttca atcgccccct      300
ggatatagcc ccgacaatag gccgtggcct catttttttg ccttccgcac atttccattg      360
ctcggtaccc acaccttgct tctcctgcac ttgccaacct taatactggt ttacattgac      420
caacatctta caagcggggg gcttgtctag ggtatatata aacagtggct ctcccaatcg      480
gttgccagtc tcttttttcc tttctttccc cacagattcg aaatctaaac tacacatcac      540
accatggaca agaaatactc catcggcctg gacattggaa ccaactctgt cggctgggct      600
gtcatcaccg acgagtacaa ggtgccctcc aagaaattca aggtcctcgg aaacaccgat      660
cgacactcca tcaagaaaaa cctcattggt gccctgttgt tcgattctgg cgagactgcc      720
gaagctacca gactcaagcg aactgctcgg cgacgttaca cccgacggaa gaaccgaatc      780
tgctacctgc aggagatctt ttccaacgag atggccaagg tggacgattc gttctttcat      840
cgactggagg aatccttcct cgtcgaggaa gacaagaaac acgagcgtca tcccatcttt      900
ggcaacattg tggacgaggt tgcttaccac gagaagtatc ctaccatcta ccacctgcga      960
aagaaactcg tcgattccac cgacaaggcg gatctcagac ttatctacct cgctctggca     1020
cacatgatca agtttcgagg tcatttcctc atcgagggcg atctcaatcc cgacaacagc     1080
gatgtggaca agctgttcat tcagctcgtt cagacctaca accagctgtt cgaggaaaac     1140
cccatcaatg cctccggagt cgatgcaaag gccatcttgt ctgctcgact ctcgaagagc     1200
agacgactgg agaacctcat tgcccaactt cctggcgaga aaaagaacgg actgtttggc     1260
aacctcattg ccctttctct tggtctcaca cccaacttca agtccaactt cgatctggcg     1320
gaggacgcca agctccagct gtccaaggac acctacgacg atgacctcga caacctgctt     1380
gcacagattg gcgatcagta cgccgacctg tttctcgctg ccaagaacct ttcggatgct     1440
attctcttgt ctgacattct gcgagtcaac accgagatca caaaggctcc cctttctgcc     1500
tccatgatca agcgatacga cgagcaccat caggatctca cactgctcaa ggctcttgtc     1560
cgacagcaac tgcccgagaa gtacaaggag atctttttcg atcagtcgaa gaacggctac     1620
gctggataca tcgacggcgg agcctctcag gaagagttct acaagttcat caagccaatt     1680
ctcgagaaga tggacggaac cgaggaactg cttgtcaagc tcaatcgaga ggatctgctt     1740
cggaagcaac gaaccttcga caacggcagc attcctcatc agatccacct cggtgagctg     1800
cacgccattc ttcgacgtca ggaagacttc tacccctttc tcaaggacaa ccgagagaag     1860
atcgagaaga ttcttacctt tcgaatcccc tactatgttg gtcctcttgc cagaggaaac     1920
tctcgatttg cttggatgac tcgaaagtcc gaggaaacca tcactccctg gaacttcgag     1980
gaagtcgtgg acaagggtgc ctctgcacag tccttcatcg agcgaatgac caacttcgac     2040
aagaatctgc ccaacgagaa ggttcttccc aagcattcgc tgctctacga gtactttaca     2100
gtctacaacg aactcaccaa agtcaagtac gttaccgagg gaatgcgaaa gcctgccttc     2160
ttgtctggcg aacagaagaa agccattgtc gatctcctgt tcaagaccaa ccgaaaggtc     2220
actgttaagc agctcaagga ggactacttc aagaaaatcg agtgtttcga cagcgtcgag     2280
atttccggag ttgaggaccg attcaacgcc tctttgggca cctatcacga tctgctcaag     2340
attatcaagg acaaggattt tctcgacaac gaggaaaacg aggacattct ggaggacatc     2400
gtgctcactc ttaccctgtt cgaagatcgg gagatgatcg aggaacgact caagacatac     2460
gctcacctgt tcgacgacaa ggtcatgaaa caactcaagc gacgtagata caccggctgg     2520
ggaagacttt cgcgaaagct catcaacggc atcagagaca agcagtccgg aaagaccatt     2580
ctggactttc tcaagtccga tggctttgcc aaccgaaact tcatgcagct cattcacgac     2640
gattctctta ccttcaagga ggacatccag aaggcacaag tgtccggtca gggcgacagc     2700
ttgcacgaac atattgccaa cctggctggt tcgccagcca tcaagaaagg cattctccag     2760
actgtcaagg ttgtcgacga gctggtgaag gtcatgggac gtcacaagcc cgagaacatt     2820
gtgatcgaga tggccagaga gaaccagaca actcaaaagg gtcagaaaaa ctcgcgagag     2880
cggatgaagc gaatcgagga aggcatcaag gagctgggat cccagattct caaggagcat     2940
cccgtcgaga acactcaact gcagaacgag aagctgtatc tctactatct gcagaatggt     3000
cgagacatgt acgtggatca ggaactggac atcaatcgtc tcagcgacta cgatgtggac     3060
cacattgtcc ctcaatcctt tctcaaggac gattctatcg acaacaaggt ccttacacga     3120
tccgacaaga acagaggcaa gtcggacaac gttcccagcg aagaggtggt caaaaagatg     3180
aagaactact ggcgacagct gctcaacgcc aagctcatta cccagcgaaa gttcgacaat     3240
cttaccaagg ccgagcgagg cggtctgtcc gagctcgaca aggctggctt catcaagcgt     3300
caactcgtcg agaccagaca gatcacaaag cacgtcgcac agattctcga ttctcggatg     3360
aacaccaagt acgacgagaa cgacaagctc atccgagagg tcaaggtgat tactctcaag     3420
tccaaactgg tctccgattt ccgaaaggac tttcagttct acaaggtgcg agagatcaac     3480
aattaccacc atgcccacga tgcttacctc aacgccgtcg ttggcactgc gctcatcaag     3540
aaatacccca agctcgaaag cgagttcgtt tacggcgatt acaaggtcta cgacgttcga     3600
aagatgattg ccaagtccga acaggagatt ggcaaggcta ctgccaagta cttcttttac     3660
tccaacatca tgaacttttt caagaccgag atcaccttgg ccaacggaga gattcgaaag     3720
agaccactta tcgagaccaa cggcgaaact ggagagatcg tgtgggacaa gggtcgagac     3780
tttgcaaccg tgcgaaaggt tctgtcgatg cctcaggtca acatcgtcaa gaaaaccgag     3840
gttcagactg gcggattctc caaggagtcg attctgccca agcgaaactc cgacaagctc     3900
atcgctcgaa agaaagactg ggatcccaag aaatacggtg gcttcgattc tcctaccgtc     3960
gcctattccg tgcttgtcgt tgcgaaggtc gagaagggca agtccaaaaa gctcaagtcc     4020
gtcaaggagc tgctcggaat taccatcatg gagcgatcga gcttcgagaa gaatcccatc     4080
gacttcttgg aagccaaggg ttacaaggag gtcaagaaag acctcattat caagctgccc     4140
aagtactctc tgttcgaact ggagaacggt cgaaagcgta tgctcgcctc cgctggcgag     4200
ctgcagaagg gaaacgagct tgccttgcct tcgaagtacg tcaactttct ctatctggct     4260
tctcactacg agaagctcaa gggttctccc gaggacaacg aacagaagca actcttcgtt     4320
gagcagcaca aacattacct cgacgagatt atcgagcaga tttccgagtt ttcgaagcga     4380
gtcatcctgg ctgatgccaa cttggacaag gtgctctctg cctacaacaa gcatcgggac     4440
aaacccattc gagaacaggc ggagaacatc attcacctgt ttactcttac caacctgggt     4500
gctcctgcag ctttcaagta cttcgatacc actatcgacc gaaagcggta cacatccacc     4560
aaggaggttc tcgatgccac cctgattcac cagtccatca ctggcctgta cgagacccga     4620
atcgacctgt ctcagcttgg tggcgactcc agagccgatc ccaagaaaaa gcgaaaggtc     4680
taa                                                                   4683

<210>  77
<211>  10706
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid  pZufCas9

<400>  77
catggacaag aaatactcca tcggcctgga cattggaacc aactctgtcg gctgggctgt       60
catcaccgac gagtacaagg tgccctccaa gaaattcaag gtcctcggaa acaccgatcg      120
acactccatc aagaaaaacc tcattggtgc cctgttgttc gattctggcg agactgccga      180
agctaccaga ctcaagcgaa ctgctcggcg acgttacacc cgacggaaga accgaatctg      240
ctacctgcag gagatctttt ccaacgagat ggccaaggtg gacgattcgt tctttcatcg      300
actggaggaa tccttcctcg tcgaggaaga caagaaacac gagcgtcatc ccatctttgg      360
caacattgtg gacgaggttg cttaccacga gaagtatcct accatctacc acctgcgaaa      420
gaaactcgtc gattccaccg acaaggcgga tctcagactt atctacctcg ctctggcaca      480
catgatcaag tttcgaggtc atttcctcat cgagggcgat ctcaatcccg acaacagcga      540
tgtggacaag ctgttcattc agctcgttca gacctacaac cagctgttcg aggaaaaccc      600
catcaatgcc tccggagtcg atgcaaaggc catcttgtct gctcgactct cgaagagcag      660
acgactggag aacctcattg cccaacttcc tggcgagaaa aagaacggac tgtttggcaa      720
cctcattgcc ctttctcttg gtctcacacc caacttcaag tccaacttcg atctggcgga      780
ggacgccaag ctccagctgt ccaaggacac ctacgacgat gacctcgaca acctgcttgc      840
acagattggc gatcagtacg ccgacctgtt tctcgctgcc aagaaccttt cggatgctat      900
tctcttgtct gacattctgc gagtcaacac cgagatcaca aaggctcccc tttctgcctc      960
catgatcaag cgatacgacg agcaccatca ggatctcaca ctgctcaagg ctcttgtccg     1020
acagcaactg cccgagaagt acaaggagat ctttttcgat cagtcgaaga acggctacgc     1080
tggatacatc gacggcggag cctctcagga agagttctac aagttcatca agccaattct     1140
cgagaagatg gacggaaccg aggaactgct tgtcaagctc aatcgagagg atctgcttcg     1200
gaagcaacga accttcgaca acggcagcat tcctcatcag atccacctcg gtgagctgca     1260
cgccattctt cgacgtcagg aagacttcta cccctttctc aaggacaacc gagagaagat     1320
cgagaagatt cttacctttc gaatccccta ctatgttggt cctcttgcca gaggaaactc     1380
tcgatttgct tggatgactc gaaagtccga ggaaaccatc actccctgga acttcgagga     1440
agtcgtggac aagggtgcct ctgcacagtc cttcatcgag cgaatgacca acttcgacaa     1500
gaatctgccc aacgagaagg ttcttcccaa gcattcgctg ctctacgagt actttacagt     1560
ctacaacgaa ctcaccaaag tcaagtacgt taccgaggga atgcgaaagc ctgccttctt     1620
gtctggcgaa cagaagaaag ccattgtcga tctcctgttc aagaccaacc gaaaggtcac     1680
tgttaagcag ctcaaggagg actacttcaa gaaaatcgag tgtttcgaca gcgtcgagat     1740
ttccggagtt gaggaccgat tcaacgcctc tttgggcacc tatcacgatc tgctcaagat     1800
tatcaaggac aaggattttc tcgacaacga ggaaaacgag gacattctgg aggacatcgt     1860
gctcactctt accctgttcg aagatcggga gatgatcgag gaacgactca agacatacgc     1920
tcacctgttc gacgacaagg tcatgaaaca actcaagcga cgtagataca ccggctgggg     1980
aagactttcg cgaaagctca tcaacggcat cagagacaag cagtccggaa agaccattct     2040
ggactttctc aagtccgatg gctttgccaa ccgaaacttc atgcagctca ttcacgacga     2100
ttctcttacc ttcaaggagg acatccagaa ggcacaagtg tccggtcagg gcgacagctt     2160
gcacgaacat attgccaacc tggctggttc gccagccatc aagaaaggca ttctccagac     2220
tgtcaaggtt gtcgacgagc tggtgaaggt catgggacgt cacaagcccg agaacattgt     2280
gatcgagatg gccagagaga accagacaac tcaaaagggt cagaaaaact cgcgagagcg     2340
gatgaagcga atcgaggaag gcatcaagga gctgggatcc cagattctca aggagcatcc     2400
cgtcgagaac actcaactgc agaacgagaa gctgtatctc tactatctgc agaatggtcg     2460
agacatgtac gtggatcagg aactggacat caatcgtctc agcgactacg atgtggacca     2520
cattgtccct caatcctttc tcaaggacga ttctatcgac aacaaggtcc ttacacgatc     2580
cgacaagaac agaggcaagt cggacaacgt tcccagcgaa gaggtggtca aaaagatgaa     2640
gaactactgg cgacagctgc tcaacgccaa gctcattacc cagcgaaagt tcgacaatct     2700
taccaaggcc gagcgaggcg gtctgtccga gctcgacaag gctggcttca tcaagcgtca     2760
actcgtcgag accagacaga tcacaaagca cgtcgcacag attctcgatt ctcggatgaa     2820
caccaagtac gacgagaacg acaagctcat ccgagaggtc aaggtgatta ctctcaagtc     2880
caaactggtc tccgatttcc gaaaggactt tcagttctac aaggtgcgag agatcaacaa     2940
ttaccaccat gcccacgatg cttacctcaa cgccgtcgtt ggcactgcgc tcatcaagaa     3000
ataccccaag ctcgaaagcg agttcgttta cggcgattac aaggtctacg acgttcgaaa     3060
gatgattgcc aagtccgaac aggagattgg caaggctact gccaagtact tcttttactc     3120
caacatcatg aactttttca agaccgagat caccttggcc aacggagaga ttcgaaagag     3180
accacttatc gagaccaacg gcgaaactgg agagatcgtg tgggacaagg gtcgagactt     3240
tgcaaccgtg cgaaaggttc tgtcgatgcc tcaggtcaac atcgtcaaga aaaccgaggt     3300
tcagactggc ggattctcca aggagtcgat tctgcccaag cgaaactccg acaagctcat     3360
cgctcgaaag aaagactggg atcccaagaa atacggtggc ttcgattctc ctaccgtcgc     3420
ctattccgtg cttgtcgttg cgaaggtcga gaagggcaag tccaaaaagc tcaagtccgt     3480
caaggagctg ctcggaatta ccatcatgga gcgatcgagc ttcgagaaga atcccatcga     3540
cttcttggaa gccaagggtt acaaggaggt caagaaagac ctcattatca agctgcccaa     3600
gtactctctg ttcgaactgg agaacggtcg aaagcgtatg ctcgcctccg ctggcgagct     3660
gcagaaggga aacgagcttg ccttgccttc gaagtacgtc aactttctct atctggcttc     3720
tcactacgag aagctcaagg gttctcccga ggacaacgaa cagaagcaac tcttcgttga     3780
gcagcacaaa cattacctcg acgagattat cgagcagatt tccgagtttt cgaagcgagt     3840
catcctggct gatgccaact tggacaaggt gctctctgcc tacaacaagc atcgggacaa     3900
acccattcga gaacaggcgg agaacatcat tcacctgttt actcttacca acctgggtgc     3960
tcctgcagct ttcaagtact tcgataccac tatcgaccga aagcggtaca catccaccaa     4020
ggaggttctc gatgccaccc tgattcacca gtccatcact ggcctgtacg agacccgaat     4080
cgacctgtct cagcttggtg gcgactccag agccgatccc aagaaaaagc gaaaggtcta     4140
agcggccgca agtgtggatg gggaagtgag tgcccggttc tgtgtgcaca attggcaatc     4200
caagatggat ggattcaaca cagggatata gcgagctacg tggtggtgcg aggatatagc     4260
aacggatatt tatgtttgac acttgagaat gtacgataca agcactgtcc aagtacaata     4320
ctaaacatac tgtacatact catactcgta cccgggcaac ggtttcactt gagtgcagtg     4380
gctagtgctc ttactcgtac agtgtgcaat actgcgtatc atagtctttg atgtatatcg     4440
tattcattca tgttagttgc gtacgagccg gaagcataaa gtgtaaagcc tggggtgcct     4500
aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa     4560
acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta     4620
ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc     4680
gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg     4740
caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt     4800
tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa     4860
gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct     4920
ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc     4980
cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg     5040
tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct     5100
tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag     5160
cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga     5220
agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga     5280
agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg     5340
gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag     5400
aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag     5460
ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat     5520
gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct     5580
taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac     5640
tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa     5700
tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg     5760
gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt     5820
gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca     5880
ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt     5940
cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct     6000
tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg     6060
cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg     6120
agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg     6180
cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa     6240
aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt     6300
aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt     6360
gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt     6420
gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca     6480
tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat     6540
ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg     6600
tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt     6660
tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc     6720
tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg     6780
gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg     6840
agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct     6900
cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg     6960
agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca     7020
ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt     7080
acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt     7140
ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg     7200
cgaattgggt accgggcccc ccctcgaggt cgatggtgtc gataagcttg atatcgaatt     7260
catgtcacac aaaccgatct tcgcctcaag gaaacctaat tctacatccg agagactgcc     7320
gagatccagt ctacactgat taattttcgg gccaataatt taaaaaaatc gtgttatata     7380
atattatatg tattatatat atacatcatg atgatactga cagtcatgtc ccattgctaa     7440
atagacagac tccatctgcc gcctccaact gatgttctca atatttaagg ggtcatctcg     7500
cattgtttaa taataaacag actccatcta ccgcctccaa atgatgttct caaaatatat     7560
tgtatgaact tatttttatt acttagtatt attagacaac ttacttgctt tatgaaaaac     7620
acttcctatt taggaaacaa tttataatgg cagttcgttc atttaacaat ttatgtagaa     7680
taaatgttat aaatgcgtat gggaaatctt aaatatggat agcataaatg atatctgcat     7740
tgcctaattc gaaatcaaca gcaacgaaaa aaatcccttg tacaacataa atagtcatcg     7800
agaaatatca actatcaaag aacagctatt cacacgttac tattgagatt attattggac     7860
gagaatcaca cactcaactg tctttctctc ttctagaaat acaggtacaa gtatgtacta     7920
ttctcattgt tcatacttct agtcatttca tcccacatat tccttggatt tctctccaat     7980
gaatgacatt ctatcttgca aattcaacaa ttataataag atataccaaa gtagcggtat     8040
agtggcaatc aaaaagcttc tctggtgtgc ttctcgtatt tatttttatt ctaatgatcc     8100
attaaaggta tatatttatt tcttgttata taatcctttt gtttattaca tgggctggat     8160
acataaaggt attttgattt aattttttgc ttaaattcaa tcccccctcg ttcagtgtca     8220
actgtaatgg taggaaatta ccatactttt gaagaagcaa aaaaaatgaa agaaaaaaaa     8280
aatcgtattt ccaggttaga cgttccgcag aatctagaat gcggtatgcg gtacattgtt     8340
cttcgaacgt aaaagttgcg ctccctgaga tattgtacat ttttgctttt acaagtacaa     8400
gtacatcgta caactatgta ctactgttga tgcatccaca acagtttgtt ttgttttttt     8460
ttgttttttt tttttctaat gattcattac cgctatgtat acctacttgt acttgtagta     8520
agccgggtta ttggcgttca attaatcata gacttatgaa tctgcacggt gtgcgctgcg     8580
agttactttt agcttatgca tgctacttgg gtgtaatatt gggatctgtt cggaaatcaa     8640
cggatgctca atcgatttcg acagtaatta attaagtcat acacaagtca gctttcttcg     8700
agcctcatat aagtataagt agttcaacgt attagcactg tacccagcat ctccgtatcg     8760
agaaacacaa caacatgccc cattggacag atcatgcgga tacacaggtt gtgcagtatc     8820
atacatactc gatcagacag gtcgtctgac catcatacaa gctgaacaag cgctccatac     8880
ttgcacgctc tctatataca cagttaaatt acatatccat agtctaacct ctaacagtta     8940
atcttctggt aagcctccca gccagccttc tggtatcgct tggcctcctc aataggatct     9000
cggttctggc cgtacagacc tcggccgaca attatgatat ccgttccggt agacatgaca     9060
tcctcaacag ttcggtactg ctgtccgaga gcgtctccct tgtcgtcaag acccaccccg     9120
ggggtcagaa taagccagtc ctcagagtcg cccttaggtc ggttctgggc aatgaagcca     9180
accacaaact cggggtcgga tcgggcaagc tcaatggtct gcttggagta ctcgccagtg     9240
gccagagagc ccttgcaaga cagctcggcc agcatgagca gacctctggc cagcttctcg     9300
ttgggagagg ggactaggaa ctccttgtac tgggagttct cgtagtcaga gacgtcctcc     9360
ttcttctgtt cagagacagt ttcctcggca ccagctcgca ggccagcaat gattccggtt     9420
ccgggtacac cgtgggcgtt ggtgatatcg gaccactcgg cgattcggtg acaccggtac     9480
tggtgcttga cagtgttgcc aatatctgcg aactttctgt cctcgaacag gaagaaaccg     9540
tgcttaagag caagttcctt gagggggagc acagtgccgg cgtaggtgaa gtcgtcaatg     9600
atgtcgatat gggttttgat catgcacaca taaggtccga ccttatcggc aagctcaatg     9660
agctccttgg tggtggtaac atccagagaa gcacacaggt tggttttctt ggctgccacg     9720
agcttgagca ctcgagcggc aaaggcggac ttgtggacgt tagctcgagc ttcgtaggag     9780
ggcattttgg tggtgaagag gagactgaaa taaatttagt ctgcagaact ttttatcgga     9840
accttatctg gggcagtgaa gtatatgtta tggtaatagt tacgagttag ttgaacttat     9900
agatagactg gactatacgg ctatcggtcc aaattagaaa gaacgtcaat ggctctctgg     9960
gcgtcgcctt tgccgacaaa aatgtgatca tgatgaaagc cagcaatgac gttgcagctg    10020
atattgttgt cggccaaccg cgccgaaaac gcagctgtca gacccacagc ctccaacgaa    10080
gaatgtatcg tcaaagtgat ccaagcacac tcatagttgg agtcgtactc caaaggcggc    10140
aatgacgagt cagacagata ctcgtcgacg tttaaaccat catctaaggg cctcaaaact    10200
acctcggaac tgctgcgctg atctggacac cacagaggtt ccgagcactt taggttgcac    10260
caaatgtccc accaggtgca ggcagaaaac gctggaacag cgtgtacagt ttgtcttaac    10320
aaaaagtgag ggcgctgagg tcgagcaggg tggtgtgact tgttatagcc tttagagctg    10380
cgaaagcgcg tatggatttg gctcatcagg ccagattgag ggtctgtgga cacatgtcat    10440
gttagtgtac ttcaatcgcc ccctggatat agccccgaca ataggccgtg gcctcatttt    10500
tttgccttcc gcacatttcc attgctcggt acccacacct tgcttctcct gcacttgcca    10560
accttaatac tggtttacat tgaccaacat cttacaagcg gggggcttgt ctagggtata    10620
tataaacagt ggctctccca atcggttgcc agtctctttt ttcctttctt tccccacaga    10680
ttcgaaatct aaactacaca tcacac                                         10706

<210>  78
<211>  4144
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cas9-SV40 fusion

<400>  78
acaagaaata ctccatcggc ctggacattg gaaccaactc tgtcggctgg gctgtcatca       60
ccgacgagta caaggtgccc tccaagaaat tcaaggtcct cggaaacacc gatcgacact      120
ccatcaagaa aaacctcatt ggtgccctgt tgttcgattc tggcgagact gccgaagcta      180
ccagactcaa gcgaactgct cggcgacgtt acacccgacg gaagaaccga atctgctacc      240
tgcaggagat cttttccaac gagatggcca aggtggacga ttcgttcttt catcgactgg      300
aggaatcctt cctcgtcgag gaagacaaga aacacgagcg tcatcccatc tttggcaaca      360
ttgtggacga ggttgcttac cacgagaagt atcctaccat ctaccacctg cgaaagaaac      420
tcgtcgattc caccgacaag gcggatctca gacttatcta cctcgctctg gcacacatga      480
tcaagtttcg aggtcatttc ctcatcgagg gcgatctcaa tcccgacaac agcgatgtgg      540
acaagctgtt cattcagctc gttcagacct acaaccagct gttcgaggaa aaccccatca      600
atgcctccgg agtcgatgca aaggccatct tgtctgctcg actctcgaag agcagacgac      660
tggagaacct cattgcccaa cttcctggcg agaaaaagaa cggactgttt ggcaacctca      720
ttgccctttc tcttggtctc acacccaact tcaagtccaa cttcgatctg gcggaggacg      780
ccaagctcca gctgtccaag gacacctacg acgatgacct cgacaacctg cttgcacaga      840
ttggcgatca gtacgccgac ctgtttctcg ctgccaagaa cctttcggat gctattctct      900
tgtctgacat tctgcgagtc aacaccgaga tcacaaaggc tcccctttct gcctccatga      960
tcaagcgata cgacgagcac catcaggatc tcacactgct caaggctctt gtccgacagc     1020
aactgcccga gaagtacaag gagatctttt tcgatcagtc gaagaacggc tacgctggat     1080
acatcgacgg cggagcctct caggaagagt tctacaagtt catcaagcca attctcgaga     1140
agatggacgg aaccgaggaa ctgcttgtca agctcaatcg agaggatctg cttcggaagc     1200
aacgaacctt cgacaacggc agcattcctc atcagatcca cctcggtgag ctgcacgcca     1260
ttcttcgacg tcaggaagac ttctacccct ttctcaagga caaccgagag aagatcgaga     1320
agattcttac ctttcgaatc ccctactatg ttggtcctct tgccagagga aactctcgat     1380
ttgcttggat gactcgaaag tccgaggaaa ccatcactcc ctggaacttc gaggaagtcg     1440
tggacaaggg tgcctctgca cagtccttca tcgagcgaat gaccaacttc gacaagaatc     1500
tgcccaacga gaaggttctt cccaagcatt cgctgctcta cgagtacttt acagtctaca     1560
acgaactcac caaagtcaag tacgttaccg agggaatgcg aaagcctgcc ttcttgtctg     1620
gcgaacagaa gaaagccatt gtcgatctcc tgttcaagac caaccgaaag gtcactgtta     1680
agcagctcaa ggaggactac ttcaagaaaa tcgagtgttt cgacagcgtc gagatttccg     1740
gagttgagga ccgattcaac gcctctttgg gcacctatca cgatctgctc aagattatca     1800
aggacaagga ttttctcgac aacgaggaaa acgaggacat tctggaggac atcgtgctca     1860
ctcttaccct gttcgaagat cgggagatga tcgaggaacg actcaagaca tacgctcacc     1920
tgttcgacga caaggtcatg aaacaactca agcgacgtag atacaccggc tggggaagac     1980
tttcgcgaaa gctcatcaac ggcatcagag acaagcagtc cggaaagacc attctggact     2040
ttctcaagtc cgatggcttt gccaaccgaa acttcatgca gctcattcac gacgattctc     2100
ttaccttcaa ggaggacatc cagaaggcac aagtgtccgg tcagggcgac agcttgcacg     2160
aacatattgc caacctggct ggttcgccag ccatcaagaa aggcattctc cagactgtca     2220
aggttgtcga cgagctggtg aaggtcatgg gacgtcacaa gcccgagaac attgtgatcg     2280
agatggccag agagaaccag acaactcaaa agggtcagaa aaactcgcga gagcggatga     2340
agcgaatcga ggaaggcatc aaggagctgg gatcccagat tctcaaggag catcccgtcg     2400
agaacactca actgcagaac gagaagctgt atctctacta tctgcagaat ggtcgagaca     2460
tgtacgtgga tcaggaactg gacatcaatc gtctcagcga ctacgatgtg gaccacattg     2520
tccctcaatc ctttctcaag gacgattcta tcgacaacaa ggtccttaca cgatccgaca     2580
agaacagagg caagtcggac aacgttccca gcgaagaggt ggtcaaaaag atgaagaact     2640
actggcgaca gctgctcaac gccaagctca ttacccagcg aaagttcgac aatcttacca     2700
aggccgagcg aggcggtctg tccgagctcg acaaggctgg cttcatcaag cgtcaactcg     2760
tcgagaccag acagatcaca aagcacgtcg cacagattct cgattctcgg atgaacacca     2820
agtacgacga gaacgacaag ctcatccgag aggtcaaggt gattactctc aagtccaaac     2880
tggtctccga tttccgaaag gactttcagt tctacaaggt gcgagagatc aacaattacc     2940
accatgccca cgatgcttac ctcaacgccg tcgttggcac tgcgctcatc aagaaatacc     3000
ccaagctcga aagcgagttc gtttacggcg attacaaggt ctacgacgtt cgaaagatga     3060
ttgccaagtc cgaacaggag attggcaagg ctactgccaa gtacttcttt tactccaaca     3120
tcatgaactt tttcaagacc gagatcacct tggccaacgg agagattcga aagagaccac     3180
ttatcgagac caacggcgaa actggagaga tcgtgtggga caagggtcga gactttgcaa     3240
ccgtgcgaaa ggttctgtcg atgcctcagg tcaacatcgt caagaaaacc gaggttcaga     3300
ctggcggatt ctccaaggag tcgattctgc ccaagcgaaa ctccgacaag ctcatcgctc     3360
gaaagaaaga ctgggatccc aagaaatacg gtggcttcga ttctcctacc gtcgcctatt     3420
ccgtgcttgt cgttgcgaag gtcgagaagg gcaagtccaa aaagctcaag tccgtcaagg     3480
agctgctcgg aattaccatc atggagcgat cgagcttcga gaagaatccc atcgacttct     3540
tggaagccaa gggttacaag gaggtcaaga aagacctcat tatcaagctg cccaagtact     3600
ctctgttcga actggagaac ggtcgaaagc gtatgctcgc ctccgctggc gagctgcaga     3660
agggaaacga gcttgccttg ccttcgaagt acgtcaactt tctctatctg gcttctcact     3720
acgagaagct caagggttct cccgaggaca acgaacagaa gcaactcttc gttgagcagc     3780
acaaacatta cctcgacgag attatcgagc agatttccga gttttcgaag cgagtcatcc     3840
tggctgatgc caacttggac aaggtgctct ctgcctacaa caagcatcgg gacaaaccca     3900
ttcgagaaca ggcggagaac atcattcacc tgtttactct taccaacctg ggtgctcctg     3960
cagctttcaa gtacttcgat accactatcg accgaaagcg gtacacatcc accaaggagg     4020
ttctcgatgc caccctgatt caccagtcca tcactggcct gtacgagacc cgaatcgacc     4080
tgtctcagct tggtggcgac tccagagccg atcccaagaa aaagcgaaag gtctaagcgg     4140
ccgc                                                                  4144

<210>  79
<211>  35
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cas9-SV40 forward primer

<400>  79
gggggaattc gacaagaaat actccatcgg cctgg                                  35

<210>  80
<211>  31
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cas9-SV40 reverse primer

<400>  80
ccccaagctt agcggccgct tagacctttc g                                      31

<210>  81
<211>  4166
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cas9-SV40 PCR product

<400>  81
gggggaattc gacaagaaat actccatcgg cctggacatt ggaaccaact ctgtcggctg       60
ggctgtcatc accgacgagt acaaggtgcc ctccaagaaa ttcaaggtcc tcggaaacac      120
cgatcgacac tccatcaaga aaaacctcat tggtgccctg ttgttcgatt ctggcgagac      180
tgccgaagct accagactca agcgaactgc tcggcgacgt tacacccgac ggaagaaccg      240
aatctgctac ctgcaggaga tcttttccaa cgagatggcc aaggtggacg attcgttctt      300
tcatcgactg gaggaatcct tcctcgtcga ggaagacaag aaacacgagc gtcatcccat      360
ctttggcaac attgtggacg aggttgctta ccacgagaag tatcctacca tctaccacct      420
gcgaaagaaa ctcgtcgatt ccaccgacaa ggcggatctc agacttatct acctcgctct      480
ggcacacatg atcaagtttc gaggtcattt cctcatcgag ggcgatctca atcccgacaa      540
cagcgatgtg gacaagctgt tcattcagct cgttcagacc tacaaccagc tgttcgagga      600
aaaccccatc aatgcctccg gagtcgatgc aaaggccatc ttgtctgctc gactctcgaa      660
gagcagacga ctggagaacc tcattgccca acttcctggc gagaaaaaga acggactgtt      720
tggcaacctc attgcccttt ctcttggtct cacacccaac ttcaagtcca acttcgatct      780
ggcggaggac gccaagctcc agctgtccaa ggacacctac gacgatgacc tcgacaacct      840
gcttgcacag attggcgatc agtacgccga cctgtttctc gctgccaaga acctttcgga      900
tgctattctc ttgtctgaca ttctgcgagt caacaccgag atcacaaagg ctcccctttc      960
tgcctccatg atcaagcgat acgacgagca ccatcaggat ctcacactgc tcaaggctct     1020
tgtccgacag caactgcccg agaagtacaa ggagatcttt ttcgatcagt cgaagaacgg     1080
ctacgctgga tacatcgacg gcggagcctc tcaggaagag ttctacaagt tcatcaagcc     1140
aattctcgag aagatggacg gaaccgagga actgcttgtc aagctcaatc gagaggatct     1200
gcttcggaag caacgaacct tcgacaacgg cagcattcct catcagatcc acctcggtga     1260
gctgcacgcc attcttcgac gtcaggaaga cttctacccc tttctcaagg acaaccgaga     1320
gaagatcgag aagattctta cctttcgaat cccctactat gttggtcctc ttgccagagg     1380
aaactctcga tttgcttgga tgactcgaaa gtccgaggaa accatcactc cctggaactt     1440
cgaggaagtc gtggacaagg gtgcctctgc acagtccttc atcgagcgaa tgaccaactt     1500
cgacaagaat ctgcccaacg agaaggttct tcccaagcat tcgctgctct acgagtactt     1560
tacagtctac aacgaactca ccaaagtcaa gtacgttacc gagggaatgc gaaagcctgc     1620
cttcttgtct ggcgaacaga agaaagccat tgtcgatctc ctgttcaaga ccaaccgaaa     1680
ggtcactgtt aagcagctca aggaggacta cttcaagaaa atcgagtgtt tcgacagcgt     1740
cgagatttcc ggagttgagg accgattcaa cgcctctttg ggcacctatc acgatctgct     1800
caagattatc aaggacaagg attttctcga caacgaggaa aacgaggaca ttctggagga     1860
catcgtgctc actcttaccc tgttcgaaga tcgggagatg atcgaggaac gactcaagac     1920
atacgctcac ctgttcgacg acaaggtcat gaaacaactc aagcgacgta gatacaccgg     1980
ctggggaaga ctttcgcgaa agctcatcaa cggcatcaga gacaagcagt ccggaaagac     2040
cattctggac tttctcaagt ccgatggctt tgccaaccga aacttcatgc agctcattca     2100
cgacgattct cttaccttca aggaggacat ccagaaggca caagtgtccg gtcagggcga     2160
cagcttgcac gaacatattg ccaacctggc tggttcgcca gccatcaaga aaggcattct     2220
ccagactgtc aaggttgtcg acgagctggt gaaggtcatg ggacgtcaca agcccgagaa     2280
cattgtgatc gagatggcca gagagaacca gacaactcaa aagggtcaga aaaactcgcg     2340
agagcggatg aagcgaatcg aggaaggcat caaggagctg ggatcccaga ttctcaagga     2400
gcatcccgtc gagaacactc aactgcagaa cgagaagctg tatctctact atctgcagaa     2460
tggtcgagac atgtacgtgg atcaggaact ggacatcaat cgtctcagcg actacgatgt     2520
ggaccacatt gtccctcaat cctttctcaa ggacgattct atcgacaaca aggtccttac     2580
acgatccgac aagaacagag gcaagtcgga caacgttccc agcgaagagg tggtcaaaaa     2640
gatgaagaac tactggcgac agctgctcaa cgccaagctc attacccagc gaaagttcga     2700
caatcttacc aaggccgagc gaggcggtct gtccgagctc gacaaggctg gcttcatcaa     2760
gcgtcaactc gtcgagacca gacagatcac aaagcacgtc gcacagattc tcgattctcg     2820
gatgaacacc aagtacgacg agaacgacaa gctcatccga gaggtcaagg tgattactct     2880
caagtccaaa ctggtctccg atttccgaaa ggactttcag ttctacaagg tgcgagagat     2940
caacaattac caccatgccc acgatgctta cctcaacgcc gtcgttggca ctgcgctcat     3000
caagaaatac cccaagctcg aaagcgagtt cgtttacggc gattacaagg tctacgacgt     3060
tcgaaagatg attgccaagt ccgaacagga gattggcaag gctactgcca agtacttctt     3120
ttactccaac atcatgaact ttttcaagac cgagatcacc ttggccaacg gagagattcg     3180
aaagagacca cttatcgaga ccaacggcga aactggagag atcgtgtggg acaagggtcg     3240
agactttgca accgtgcgaa aggttctgtc gatgcctcag gtcaacatcg tcaagaaaac     3300
cgaggttcag actggcggat tctccaagga gtcgattctg cccaagcgaa actccgacaa     3360
gctcatcgct cgaaagaaag actgggatcc caagaaatac ggtggcttcg attctcctac     3420
cgtcgcctat tccgtgcttg tcgttgcgaa ggtcgagaag ggcaagtcca aaaagctcaa     3480
gtccgtcaag gagctgctcg gaattaccat catggagcga tcgagcttcg agaagaatcc     3540
catcgacttc ttggaagcca agggttacaa ggaggtcaag aaagacctca ttatcaagct     3600
gcccaagtac tctctgttcg aactggagaa cggtcgaaag cgtatgctcg cctccgctgg     3660
cgagctgcag aagggaaacg agcttgcctt gccttcgaag tacgtcaact ttctctatct     3720
ggcttctcac tacgagaagc tcaagggttc tcccgaggac aacgaacaga agcaactctt     3780
cgttgagcag cacaaacatt acctcgacga gattatcgag cagatttccg agttttcgaa     3840
gcgagtcatc ctggctgatg ccaacttgga caaggtgctc tctgcctaca acaagcatcg     3900
ggacaaaccc attcgagaac aggcggagaa catcattcac ctgtttactc ttaccaacct     3960
gggtgctcct gcagctttca agtacttcga taccactatc gaccgaaagc ggtacacatc     4020
caccaaggag gttctcgatg ccaccctgat tcaccagtcc atcactggcc tgtacgagac     4080
ccgaatcgac ctgtctcagc ttggtggcga ctccagagcc gatcccaaga aaaagcgaaa     4140
ggtctaagcg gccgctaagc ttgggg                                          4166

<210>  82
<211>  4092
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pBAD/HisB

<400>  82
aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct       60
tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca      120
aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg      180
attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg      240
atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc      300
taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc      360
atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc      420
tcgagatctg cagctggtac catatgggaa ttcgaagctt ggctgttttg gcggatgaga      480
gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa      540
tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa      600
acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc      660
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt      720
cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc      780
aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc      840
agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc      900
taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa      960
tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt     1020
gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct     1080
gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc     1140
cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta     1200
tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac     1260
tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc     1320
atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac     1380
ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg     1440
gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac     1500
gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc     1560
gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt     1620
gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga     1680
gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc     1740
cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag     1800
atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca     1860
tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc     1920
ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca     1980
gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc     2040
tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta     2100
ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt     2160
ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc     2220
gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg     2280
ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg     2340
tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag     2400
ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc     2460
agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat     2520
agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg     2580
gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc     2640
tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt     2700
accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca     2760
gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt     2820
atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc     2880
cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa     2940
cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg     3000
tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga     3060
ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg     3120
acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg     3180
ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga     3240
actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt     3300
caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct     3360
tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc     3420
ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat     3480
caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat     3540
ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa     3600
gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt     3660
gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat     3720
tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa     3780
cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg     3840
gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca     3900
ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga     3960
taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat     4020
taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc     4080
cgccattcag ag                                                         4092

<210>  83
<211>  8237
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plamsid pRF48

<400>  83
aattcgacaa gaaatactcc atcggcctgg acattggaac caactctgtc ggctgggctg       60
tcatcaccga cgagtacaag gtgccctcca agaaattcaa ggtcctcgga aacaccgatc      120
gacactccat caagaaaaac ctcattggtg ccctgttgtt cgattctggc gagactgccg      180
aagctaccag actcaagcga actgctcggc gacgttacac ccgacggaag aaccgaatct      240
gctacctgca ggagatcttt tccaacgaga tggccaaggt ggacgattcg ttctttcatc      300
gactggagga atccttcctc gtcgaggaag acaagaaaca cgagcgtcat cccatctttg      360
gcaacattgt ggacgaggtt gcttaccacg agaagtatcc taccatctac cacctgcgaa      420
agaaactcgt cgattccacc gacaaggcgg atctcagact tatctacctc gctctggcac      480
acatgatcaa gtttcgaggt catttcctca tcgagggcga tctcaatccc gacaacagcg      540
atgtggacaa gctgttcatt cagctcgttc agacctacaa ccagctgttc gaggaaaacc      600
ccatcaatgc ctccggagtc gatgcaaagg ccatcttgtc tgctcgactc tcgaagagca      660
gacgactgga gaacctcatt gcccaacttc ctggcgagaa aaagaacgga ctgtttggca      720
acctcattgc cctttctctt ggtctcacac ccaacttcaa gtccaacttc gatctggcgg      780
aggacgccaa gctccagctg tccaaggaca cctacgacga tgacctcgac aacctgcttg      840
cacagattgg cgatcagtac gccgacctgt ttctcgctgc caagaacctt tcggatgcta      900
ttctcttgtc tgacattctg cgagtcaaca ccgagatcac aaaggctccc ctttctgcct      960
ccatgatcaa gcgatacgac gagcaccatc aggatctcac actgctcaag gctcttgtcc     1020
gacagcaact gcccgagaag tacaaggaga tctttttcga tcagtcgaag aacggctacg     1080
ctggatacat cgacggcgga gcctctcagg aagagttcta caagttcatc aagccaattc     1140
tcgagaagat ggacggaacc gaggaactgc ttgtcaagct caatcgagag gatctgcttc     1200
ggaagcaacg aaccttcgac aacggcagca ttcctcatca gatccacctc ggtgagctgc     1260
acgccattct tcgacgtcag gaagacttct acccctttct caaggacaac cgagagaaga     1320
tcgagaagat tcttaccttt cgaatcccct actatgttgg tcctcttgcc agaggaaact     1380
ctcgatttgc ttggatgact cgaaagtccg aggaaaccat cactccctgg aacttcgagg     1440
aagtcgtgga caagggtgcc tctgcacagt ccttcatcga gcgaatgacc aacttcgaca     1500
agaatctgcc caacgagaag gttcttccca agcattcgct gctctacgag tactttacag     1560
tctacaacga actcaccaaa gtcaagtacg ttaccgaggg aatgcgaaag cctgccttct     1620
tgtctggcga acagaagaaa gccattgtcg atctcctgtt caagaccaac cgaaaggtca     1680
ctgttaagca gctcaaggag gactacttca agaaaatcga gtgtttcgac agcgtcgaga     1740
tttccggagt tgaggaccga ttcaacgcct ctttgggcac ctatcacgat ctgctcaaga     1800
ttatcaagga caaggatttt ctcgacaacg aggaaaacga ggacattctg gaggacatcg     1860
tgctcactct taccctgttc gaagatcggg agatgatcga ggaacgactc aagacatacg     1920
ctcacctgtt cgacgacaag gtcatgaaac aactcaagcg acgtagatac accggctggg     1980
gaagactttc gcgaaagctc atcaacggca tcagagacaa gcagtccgga aagaccattc     2040
tggactttct caagtccgat ggctttgcca accgaaactt catgcagctc attcacgacg     2100
attctcttac cttcaaggag gacatccaga aggcacaagt gtccggtcag ggcgacagct     2160
tgcacgaaca tattgccaac ctggctggtt cgccagccat caagaaaggc attctccaga     2220
ctgtcaaggt tgtcgacgag ctggtgaagg tcatgggacg tcacaagccc gagaacattg     2280
tgatcgagat ggccagagag aaccagacaa ctcaaaaggg tcagaaaaac tcgcgagagc     2340
ggatgaagcg aatcgaggaa ggcatcaagg agctgggatc ccagattctc aaggagcatc     2400
ccgtcgagaa cactcaactg cagaacgaga agctgtatct ctactatctg cagaatggtc     2460
gagacatgta cgtggatcag gaactggaca tcaatcgtct cagcgactac gatgtggacc     2520
acattgtccc tcaatccttt ctcaaggacg attctatcga caacaaggtc cttacacgat     2580
ccgacaagaa cagaggcaag tcggacaacg ttcccagcga agaggtggtc aaaaagatga     2640
agaactactg gcgacagctg ctcaacgcca agctcattac ccagcgaaag ttcgacaatc     2700
ttaccaaggc cgagcgaggc ggtctgtccg agctcgacaa ggctggcttc atcaagcgtc     2760
aactcgtcga gaccagacag atcacaaagc acgtcgcaca gattctcgat tctcggatga     2820
acaccaagta cgacgagaac gacaagctca tccgagaggt caaggtgatt actctcaagt     2880
ccaaactggt ctccgatttc cgaaaggact ttcagttcta caaggtgcga gagatcaaca     2940
attaccacca tgcccacgat gcttacctca acgccgtcgt tggcactgcg ctcatcaaga     3000
aataccccaa gctcgaaagc gagttcgttt acggcgatta caaggtctac gacgttcgaa     3060
agatgattgc caagtccgaa caggagattg gcaaggctac tgccaagtac ttcttttact     3120
ccaacatcat gaactttttc aagaccgaga tcaccttggc caacggagag attcgaaaga     3180
gaccacttat cgagaccaac ggcgaaactg gagagatcgt gtgggacaag ggtcgagact     3240
ttgcaaccgt gcgaaaggtt ctgtcgatgc ctcaggtcaa catcgtcaag aaaaccgagg     3300
ttcagactgg cggattctcc aaggagtcga ttctgcccaa gcgaaactcc gacaagctca     3360
tcgctcgaaa gaaagactgg gatcccaaga aatacggtgg cttcgattct cctaccgtcg     3420
cctattccgt gcttgtcgtt gcgaaggtcg agaagggcaa gtccaaaaag ctcaagtccg     3480
tcaaggagct gctcggaatt accatcatgg agcgatcgag cttcgagaag aatcccatcg     3540
acttcttgga agccaagggt tacaaggagg tcaagaaaga cctcattatc aagctgccca     3600
agtactctct gttcgaactg gagaacggtc gaaagcgtat gctcgcctcc gctggcgagc     3660
tgcagaaggg aaacgagctt gccttgcctt cgaagtacgt caactttctc tatctggctt     3720
ctcactacga gaagctcaag ggttctcccg aggacaacga acagaagcaa ctcttcgttg     3780
agcagcacaa acattacctc gacgagatta tcgagcagat ttccgagttt tcgaagcgag     3840
tcatcctggc tgatgccaac ttggacaagg tgctctctgc ctacaacaag catcgggaca     3900
aacccattcg agaacaggcg gagaacatca ttcacctgtt tactcttacc aacctgggtg     3960
ctcctgcagc tttcaagtac ttcgatacca ctatcgaccg aaagcggtac acatccacca     4020
aggaggttct cgatgccacc ctgattcacc agtccatcac tggcctgtac gagacccgaa     4080
tcgacctgtc tcagcttggt ggcgactcca gagccgatcc caagaaaaag cgaaaggtct     4140
aagcggccgc taagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga     4200
ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg     4260
tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg     4320
tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag     4380
tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg     4440
acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca     4500
ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc     4560
ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc     4620
gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag     4680
tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt     4740
tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt     4800
gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga     4860
acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt     4920
tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga     4980
gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag     5040
tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg     5100
accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg     5160
ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt     5220
agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg     5280
gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc     5340
ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg     5400
tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac     5460
ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact     5520
gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa     5580
acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa     5640
aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg     5700
atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc     5760
gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac     5820
tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca     5880
ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt     5940
ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc     6000
ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg     6060
aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc     6120
cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac     6180
gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct     6240
ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc     6300
cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt     6360
tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac     6420
cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg     6480
cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac     6540
tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta     6600
cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg     6660
gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg     6720
tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg     6780
aaggcgaagc ggcatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc     6840
ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg     6900
gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg     6960
cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg     7020
gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc     7080
tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac     7140
ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga     7200
tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg     7260
ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc     7320
gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc     7380
ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta     7440
agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga     7500
gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc     7560
ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga     7620
ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg     7680
gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg     7740
ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat     7800
tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc     7860
aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca     7920
aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac     7980
ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga     8040
cgctttttat cgcaactctc tactgtttct ccatacccgt tttttgggct aacaggagga     8100
attaaccatg gggggttctc atcatcatca tcatcatggt atggctagca tgactggtgg     8160
acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatccgagct cgagatctgc     8220
agctggtacc atatggg                                                    8237

<210>  84
<211>  6493
<212>  DNA
<213>  Artificial sequence

<220>
<223>  E.Coli Cas9 expression cassette

<400>  84
tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat       60
cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct      120
gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct      180
gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc      240
gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc      300
aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact      360
tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc      420
cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac      480
cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt      540
ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac      600
gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca      660
ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg      720
actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca      780
gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga      840
aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc      900
agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt      960
cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat     1020
cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg     1080
tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac     1140
cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca     1200
gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa     1260
ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc     1320
taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca     1380
tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt     1440
tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta     1500
cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt gggctaacag     1560
gaggaattaa ccatgggggg ttctcatcat catcatcatc atggtatggc tagcatgact     1620
ggtggacagc aaatgggtcg ggatctgtac gacgatgacg ataaggatcc gagctcgaga     1680
tctgcagctg gtaccatatg ggaattcgac aagaaatact ccatcggcct ggacattgga     1740
accaactctg tcggctgggc tgtcatcacc gacgagtaca aggtgccctc caagaaattc     1800
aaggtcctcg gaaacaccga tcgacactcc atcaagaaaa acctcattgg tgccctgttg     1860
ttcgattctg gcgagactgc cgaagctacc agactcaagc gaactgctcg gcgacgttac     1920
acccgacgga agaaccgaat ctgctacctg caggagatct tttccaacga gatggccaag     1980
gtggacgatt cgttctttca tcgactggag gaatccttcc tcgtcgagga agacaagaaa     2040
cacgagcgtc atcccatctt tggcaacatt gtggacgagg ttgcttacca cgagaagtat     2100
cctaccatct accacctgcg aaagaaactc gtcgattcca ccgacaaggc ggatctcaga     2160
cttatctacc tcgctctggc acacatgatc aagtttcgag gtcatttcct catcgagggc     2220
gatctcaatc ccgacaacag cgatgtggac aagctgttca ttcagctcgt tcagacctac     2280
aaccagctgt tcgaggaaaa ccccatcaat gcctccggag tcgatgcaaa ggccatcttg     2340
tctgctcgac tctcgaagag cagacgactg gagaacctca ttgcccaact tcctggcgag     2400
aaaaagaacg gactgtttgg caacctcatt gccctttctc ttggtctcac acccaacttc     2460
aagtccaact tcgatctggc ggaggacgcc aagctccagc tgtccaagga cacctacgac     2520
gatgacctcg acaacctgct tgcacagatt ggcgatcagt acgccgacct gtttctcgct     2580
gccaagaacc tttcggatgc tattctcttg tctgacattc tgcgagtcaa caccgagatc     2640
acaaaggctc ccctttctgc ctccatgatc aagcgatacg acgagcacca tcaggatctc     2700
acactgctca aggctcttgt ccgacagcaa ctgcccgaga agtacaagga gatctttttc     2760
gatcagtcga agaacggcta cgctggatac atcgacggcg gagcctctca ggaagagttc     2820
tacaagttca tcaagccaat tctcgagaag atggacggaa ccgaggaact gcttgtcaag     2880
ctcaatcgag aggatctgct tcggaagcaa cgaaccttcg acaacggcag cattcctcat     2940
cagatccacc tcggtgagct gcacgccatt cttcgacgtc aggaagactt ctaccccttt     3000
ctcaaggaca accgagagaa gatcgagaag attcttacct ttcgaatccc ctactatgtt     3060
ggtcctcttg ccagaggaaa ctctcgattt gcttggatga ctcgaaagtc cgaggaaacc     3120
atcactccct ggaacttcga ggaagtcgtg gacaagggtg cctctgcaca gtccttcatc     3180
gagcgaatga ccaacttcga caagaatctg cccaacgaga aggttcttcc caagcattcg     3240
ctgctctacg agtactttac agtctacaac gaactcacca aagtcaagta cgttaccgag     3300
ggaatgcgaa agcctgcctt cttgtctggc gaacagaaga aagccattgt cgatctcctg     3360
ttcaagacca accgaaaggt cactgttaag cagctcaagg aggactactt caagaaaatc     3420
gagtgtttcg acagcgtcga gatttccgga gttgaggacc gattcaacgc ctctttgggc     3480
acctatcacg atctgctcaa gattatcaag gacaaggatt ttctcgacaa cgaggaaaac     3540
gaggacattc tggaggacat cgtgctcact cttaccctgt tcgaagatcg ggagatgatc     3600
gaggaacgac tcaagacata cgctcacctg ttcgacgaca aggtcatgaa acaactcaag     3660
cgacgtagat acaccggctg gggaagactt tcgcgaaagc tcatcaacgg catcagagac     3720
aagcagtccg gaaagaccat tctggacttt ctcaagtccg atggctttgc caaccgaaac     3780
ttcatgcagc tcattcacga cgattctctt accttcaagg aggacatcca gaaggcacaa     3840
gtgtccggtc agggcgacag cttgcacgaa catattgcca acctggctgg ttcgccagcc     3900
atcaagaaag gcattctcca gactgtcaag gttgtcgacg agctggtgaa ggtcatggga     3960
cgtcacaagc ccgagaacat tgtgatcgag atggccagag agaaccagac aactcaaaag     4020
ggtcagaaaa actcgcgaga gcggatgaag cgaatcgagg aaggcatcaa ggagctggga     4080
tcccagattc tcaaggagca tcccgtcgag aacactcaac tgcagaacga gaagctgtat     4140
ctctactatc tgcagaatgg tcgagacatg tacgtggatc aggaactgga catcaatcgt     4200
ctcagcgact acgatgtgga ccacattgtc cctcaatcct ttctcaagga cgattctatc     4260
gacaacaagg tccttacacg atccgacaag aacagaggca agtcggacaa cgttcccagc     4320
gaagaggtgg tcaaaaagat gaagaactac tggcgacagc tgctcaacgc caagctcatt     4380
acccagcgaa agttcgacaa tcttaccaag gccgagcgag gcggtctgtc cgagctcgac     4440
aaggctggct tcatcaagcg tcaactcgtc gagaccagac agatcacaaa gcacgtcgca     4500
cagattctcg attctcggat gaacaccaag tacgacgaga acgacaagct catccgagag     4560
gtcaaggtga ttactctcaa gtccaaactg gtctccgatt tccgaaagga ctttcagttc     4620
tacaaggtgc gagagatcaa caattaccac catgcccacg atgcttacct caacgccgtc     4680
gttggcactg cgctcatcaa gaaatacccc aagctcgaaa gcgagttcgt ttacggcgat     4740
tacaaggtct acgacgttcg aaagatgatt gccaagtccg aacaggagat tggcaaggct     4800
actgccaagt acttctttta ctccaacatc atgaactttt tcaagaccga gatcaccttg     4860
gccaacggag agattcgaaa gagaccactt atcgagacca acggcgaaac tggagagatc     4920
gtgtgggaca agggtcgaga ctttgcaacc gtgcgaaagg ttctgtcgat gcctcaggtc     4980
aacatcgtca agaaaaccga ggttcagact ggcggattct ccaaggagtc gattctgccc     5040
aagcgaaact ccgacaagct catcgctcga aagaaagact gggatcccaa gaaatacggt     5100
ggcttcgatt ctcctaccgt cgcctattcc gtgcttgtcg ttgcgaaggt cgagaagggc     5160
aagtccaaaa agctcaagtc cgtcaaggag ctgctcggaa ttaccatcat ggagcgatcg     5220
agcttcgaga agaatcccat cgacttcttg gaagccaagg gttacaagga ggtcaagaaa     5280
gacctcatta tcaagctgcc caagtactct ctgttcgaac tggagaacgg tcgaaagcgt     5340
atgctcgcct ccgctggcga gctgcagaag ggaaacgagc ttgccttgcc ttcgaagtac     5400
gtcaactttc tctatctggc ttctcactac gagaagctca agggttctcc cgaggacaac     5460
gaacagaagc aactcttcgt tgagcagcac aaacattacc tcgacgagat tatcgagcag     5520
atttccgagt tttcgaagcg agtcatcctg gctgatgcca acttggacaa ggtgctctct     5580
gcctacaaca agcatcggga caaacccatt cgagaacagg cggagaacat cattcacctg     5640
tttactctta ccaacctggg tgctcctgca gctttcaagt acttcgatac cactatcgac     5700
cgaaagcggt acacatccac caaggaggtt ctcgatgcca ccctgattca ccagtccatc     5760
actggcctgt acgagacccg aatcgacctg tctcagcttg gtggcgactc cagagccgat     5820
cccaagaaaa agcgaaaggt ctaagcggcc gctaagcttg gctgttttgg cggatgagag     5880
aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat     5940
ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa     6000
cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca     6060
tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc     6120
ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca     6180
acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca     6240
gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct     6300
aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat     6360
attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg     6420
cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg     6480
aagatcagtt ggg                                                        6493

<210>  85
<211>  5681
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pKO3

<220>
<221>  misc_feature
<222>  (1039)..(1039)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (1109)..(1109)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (1661)..(1662)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (1665)..(1665)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (3573)..(3574)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (5648)..(5648)
<223>  n is a, c, g, or t

<400>  85
ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc       60
tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga      120
tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt      180
ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg      240
atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc      300
agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac      360
cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat      420
gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag      480
tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag      540
tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta      600
cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc      660
caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt      720
cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct      780
ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga      840
attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg      900
gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtttant cggtacccgg ggatcgcggc cgcggaccgg atcctctaga     1080
gcggccgcga tcctctagag tcgaccggng aatggcgaat gggacgcgcc ctgtagcggc     1140
gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc     1200
ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc     1260
cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc     1320
gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg     1380
gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact     1440
ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt     1500
tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa     1560
atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt     1620
gtttattttt ctaaatacat tcaaatatgt atccgctcat nncangatcc tttttaaccc     1680
atcacatata cctgccgttc actattattt agtgaaatga gatattatga tattttctga     1740
attgtgatta aaaaggcaac tttatgccca tgcaacagaa actataaaaa atacagagaa     1800
tgaaaagaaa cagatagatt ttttagttct ttaggcccgt agtctgcaaa tccttttatg     1860
attttctatc aaacaaaaga ggaaaataga ccagttgcaa tccaaacgag agtctaatag     1920
aatgaggtcg aaaagtaaat cgcgcgggtt tgttactgat aaagcaggca agacctaaaa     1980
tgtgtaaagg gcaaagtgta tactttggcg tcacccctta catattttag gtcttttttt     2040
attgtgcgta actaacttgc catcttcaaa caggagggct ggaagaagca gaccgctaac     2100
acagtacata aaaaaggaga catgaacgat gaacatcaaa aagtttgcaa aacaagcaac     2160
agtattaacc tttactaccg cactgctggc aggaggcgca actcaagcgt ttgcgaaaga     2220
aacgaaccaa aagccatata aggaaacata cggcatttcc catattacac gccatgatat     2280
gctgcaaatc cctgaacagc aaaaaaatga aaaatatcaa gttcctgaat tcgattcgtc     2340
cacaattaaa aatatctctt ctgcaaaagg cctggacgtt tgggacagct ggccattaca     2400
aaacgctgac ggcactgtcg caaactatca cggctaccac atcgtctttg cattagccgg     2460
agatcctaaa aatgcggatg acacatcgat ttacatgttc tatcaaaaag tcggcgaaac     2520
ttctattgac agctggaaaa acgctggccg cgtctttaaa gacagcgaca aattcgatgc     2580
aaatgattct atcctaaaag accaaacaca agaatggtca ggttcagcca catttacatc     2640
tgacggaaaa atccgtttat tctacactga tttctccggt aaacattacg gcaaacaaac     2700
actgacaact gcacaagtta acgtatcagc atcagacagc tctttgaaca tcaacggtgt     2760
agaggattat aaatcaatct ttgacggtga cggaaaaacg tatcaaaatg tacagcagtt     2820
catcgatgaa ggcaactaca gctcaggcga caaccatacg ctgagagatc ctcactacgt     2880
agaagataaa ggccacaaat acttagtatt tgaagcaaac actggaactg aagatggcta     2940
ccaaggcgaa gaatctttat ttaacaaagc atactatggc aaaagcacat cattcttccg     3000
tcaagaaagt caaaaacttc tgcaaagcga taaaaaacgc acggctgagt tagcaaacgg     3060
cgctctcggt atgattgagc taaacgatga ttacacactg aaaaaagtga tgaaaccgct     3120
gattgcatct aacacagtaa cagatgaaat tgaacgcgcg aacgtcttta aaatgaacgg     3180
caaatggtac ctgttcactg actcccgcgg atcaaaaatg acgattgacg gcattacgtc     3240
taacgatatt tacatgcttg gttatgtttc taattcttta actggcccat acaagccgct     3300
gaacaaaact ggccttgtgt taaaaatgga tcttgatcct aacgatgtaa cctttactta     3360
ctcacacttc gctgtacctc aagcgaaagg aaacaatgtc gtgattacaa gctatatgac     3420
aaacagagga ttctacgcag acaaacaatc aacgtttgcg ccaagcttcc tgctgaacat     3480
caaaggcaag aaaacatctg ttgtcaaaga cagcatcctt gaacaaggac aattaacagt     3540
taacaaataa aaacgcaaaa gaaaatgccg atnnccggtt tattgactac cggaagcagt     3600
gtgaccgtgt gcttctcaaa tgcctcaggc tgtctatgtg tgactgttga gctgtaacaa     3660
gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc     3720
tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt     3780
taaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat     3840
ctgtgcatat ggacagtttt ccctttgata tctaacggtg aacagttgtt ctacttttgt     3900
ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg     3960
tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa     4020
ccattgagat catgcttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc     4080
tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttacg taggtaggaa     4140
tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt     4200
tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg     4260
cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt     4320
ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc aagcattaac     4380
atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt     4440
agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca     4500
tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca     4560
ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc     4620
tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt     4680
gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg     4740
ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt     4800
agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc     4860
gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga     4920
ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt     4980
tgagttgtgg gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct     5040
agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt     5100
ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg     5160
tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt     5220
gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg     5280
ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt     5340
tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta     5400
caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac     5460
gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt     5520
tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt     5580
cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct     5640
tactgtcngg atcgacgctc tcccttatgc gactcctgca t                         5681

<210>  86
<211>  12166
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF97

<400>  86
ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc       60
tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga      120
tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt      180
ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg      240
atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc      300
agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac      360
cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat      420
gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag      480
tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag      540
tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta      600
cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc      660
caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt      720
cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct      780
ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga      840
attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg      900
gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtttatc ggtacccccc aactgatctt cagcatcttt tactttcacc     1080
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg     1140
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag     1200
ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt     1260
ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc     1320
agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc     1380
cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg     1440
aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc     1500
tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca     1560
tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc     1620
ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc     1680
caagcttagc ggccgcttag acctttcgct ttttcttggg atcggctctg gagtcgccac     1740
caagctgaga caggtcgatt cgggtctcgt acaggccagt gatggactgg tgaatcaggg     1800
tggcatcgag aacctccttg gtggatgtgt accgctttcg gtcgatagtg gtatcgaagt     1860
acttgaaagc tgcaggagca cccaggttgg taagagtaaa caggtgaatg atgttctccg     1920
cctgttctcg aatgggtttg tcccgatgct tgttgtaggc agagagcacc ttgtccaagt     1980
tggcatcagc caggatgact cgcttcgaaa actcggaaat ctgctcgata atctcgtcga     2040
ggtaatgttt gtgctgctca acgaagagtt gcttctgttc gttgtcctcg ggagaaccct     2100
tgagcttctc gtagtgagaa gccagataga gaaagttgac gtacttcgaa ggcaaggcaa     2160
gctcgtttcc cttctgcagc tcgccagcgg aggcgagcat acgctttcga ccgttctcca     2220
gttcgaacag agagtacttg ggcagcttga taatgaggtc tttcttgacc tccttgtaac     2280
ccttggcttc caagaagtcg atgggattct tctcgaagct cgatcgctcc atgatggtaa     2340
ttccgagcag ctccttgacg gacttgagct ttttggactt gcccttctcg accttcgcaa     2400
cgacaagcac ggaataggcg acggtaggag aatcgaagcc accgtatttc ttgggatccc     2460
agtctttctt tcgagcgatg agcttgtcgg agtttcgctt gggcagaatc gactccttgg     2520
agaatccgcc agtctgaacc tcggttttct tgacgatgtt gacctgaggc atcgacagaa     2580
cctttcgcac ggttgcaaag tctcgaccct tgtcccacac gatctctcca gtttcgccgt     2640
tggtctcgat aagtggtctc tttcgaatct ctccgttggc caaggtgatc tcggtcttga     2700
aaaagttcat gatgttggag taaaagaagt acttggcagt agccttgcca atctcctgtt     2760
cggacttggc aatcatcttt cgaacgtcgt agaccttgta atcgccgtaa acgaactcgc     2820
tttcgagctt ggggtatttc ttgatgagcg cagtgccaac gacggcgttg aggtaagcat     2880
cgtgggcatg gtggtaattg ttgatctctc gcaccttgta gaactgaaag tcctttcgga     2940
aatcggagac cagtttggac ttgagagtaa tcaccttgac ctctcggatg agcttgtcgt     3000
tctcgtcgta cttggtgttc atccgagaat cgagaatctg tgcgacgtgc tttgtgatct     3060
gtctggtctc gacgagttga cgcttgatga agccagcctt gtcgagctcg gacagaccgc     3120
ctcgctcggc cttggtaaga ttgtcgaact ttcgctgggt aatgagcttg gcgttgagca     3180
gctgtcgcca gtagttcttc atctttttga ccacctcttc gctgggaacg ttgtccgact     3240
tgcctctgtt cttgtcggat cgtgtaagga ccttgttgtc gatagaatcg tccttgagaa     3300
aggattgagg gacaatgtgg tccacatcgt agtcgctgag acgattgatg tccagttcct     3360
gatccacgta catgtctcga ccattctgca gatagtagag atacagcttc tcgttctgca     3420
gttgagtgtt ctcgacggga tgctccttga gaatctggga tcccagctcc ttgatgcctt     3480
cctcgattcg cttcatccgc tctcgcgagt ttttctgacc cttttgagtt gtctggttct     3540
ctctggccat ctcgatcaca atgttctcgg gcttgtgacg tcccatgacc ttcaccagct     3600
cgtcgacaac cttgacagtc tggagaatgc ctttcttgat ggctggcgaa ccagccaggt     3660
tggcaatatg ttcgtgcaag ctgtcgccct gaccggacac ttgtgccttc tggatgtcct     3720
ccttgaaggt aagagaatcg tcgtgaatga gctgcatgaa gtttcggttg gcaaagccat     3780
cggacttgag aaagtccaga atggtctttc cggactgctt gtctctgatg ccgttgatga     3840
gctttcgcga aagtcttccc cagccggtgt atctacgtcg cttgagttgt ttcatgacct     3900
tgtcgtcgaa caggtgagcg tatgtcttga gtcgttcctc gatcatctcc cgatcttcga     3960
acagggtaag agtgagcacg atgtcctcca gaatgtcctc gttttcctcg ttgtcgagaa     4020
aatccttgtc cttgataatc ttgagcagat cgtgataggt gcccaaagag gcgttgaatc     4080
ggtcctcaac tccggaaatc tcgacgctgt cgaaacactc gattttcttg aagtagtcct     4140
ccttgagctg cttaacagtg acctttcggt tggtcttgaa caggagatcg acaatggctt     4200
tcttctgttc gccagacaag aaggcaggct ttcgcattcc ctcggtaacg tacttgactt     4260
tggtgagttc gttgtagact gtaaagtact cgtagagcag cgaatgcttg ggaagaacct     4320
tctcgttggg cagattcttg tcgaagttgg tcattcgctc gatgaaggac tgtgcagagg     4380
cacccttgtc cacgacttcc tcgaagttcc agggagtgat ggtttcctcg gactttcgag     4440
tcatccaagc aaatcgagag tttcctctgg caagaggacc aacatagtag gggattcgaa     4500
aggtaagaat cttctcgatc ttctctcggt tgtccttgag aaaggggtag aagtcttcct     4560
gacgtcgaag aatggcgtgc agctcaccga ggtggatctg atgaggaatg ctgccgttgt     4620
cgaaggttcg ttgcttccga agcagatcct ctcgattgag cttgacaagc agttcctcgg     4680
ttccgtccat cttctcgaga attggcttga tgaacttgta gaactcttcc tgagaggctc     4740
cgccgtcgat gtatccagcg tagccgttct tcgactgatc gaaaaagatc tccttgtact     4800
tctcgggcag ttgctgtcgg acaagagcct tgagcagtgt gagatcctga tggtgctcgt     4860
cgtatcgctt gatcatggag gcagaaaggg gagcctttgt gatctcggtg ttgactcgca     4920
gaatgtcaga caagagaata gcatccgaaa ggttcttggc agcgagaaac aggtcggcgt     4980
actgatcgcc aatctgtgca agcaggttgt cgaggtcatc gtcgtaggtg tccttggaca     5040
gctggagctt ggcgtcctcc gccagatcga agttggactt gaagttgggt gtgagaccaa     5100
gagaaagggc aatgaggttg ccaaacagtc cgttcttttt ctcgccagga agttgggcaa     5160
tgaggttctc cagtcgtctg ctcttcgaga gtcgagcaga caagatggcc tttgcatcga     5220
ctccggaggc attgatgggg ttttcctcga acagctggtt gtaggtctga acgagctgaa     5280
tgaacagctt gtccacatcg ctgttgtcgg gattgagatc gccctcgatg aggaaatgac     5340
ctcgaaactt gatcatgtgt gccagagcga ggtagataag tctgagatcc gccttgtcgg     5400
tggaatcgac gagtttcttt cgcaggtggt agatggtagg atacttctcg tggtaagcaa     5460
cctcgtccac aatgttgcca aagatgggat gacgctcgtg tttcttgtct tcctcgacga     5520
ggaaggattc ctccagtcga tgaaagaacg aatcgtccac cttggccatc tcgttggaaa     5580
agatctcctg caggtagcag attcggttct tccgtcgggt gtaacgtcgc cgagcagttc     5640
gcttgagtct ggtagcttcg gcagtctcgc cagaatcgaa caacagggca ccaatgaggt     5700
ttttcttgat ggagtgtcga tcggtgtttc cgaggacctt gaatttcttg gagggcacct     5760
tgtactcgtc ggtgatgaca gcccagccga cagagttggt tccaatgtcc aggccgatgg     5820
agtatttctt gtcgaattcc catatggtac cagctgcaga tctcgagctc ggatccttat     5880
cgtcatcgtc gtacagatcc cgacccattt gctgtccacc agtcatgcta gccataccat     5940
gatgatgatg atgatgagaa ccccccatgg ttaattcctc ctgttagccc aaaaaacggg     6000
tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg     6060
gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt     6120
ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt     6180
tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag     6240
acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg     6300
agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat     6360
gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc     6420
gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg     6480
gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca     6540
ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt     6600
tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat     6660
acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa     6720
atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt     6780
gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat     6840
aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat     6900
atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc     6960
cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag     7020
ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac     7080
gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc     7140
cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg     7200
aatcagacaa ttgacggctt gacggagtag catagggttt gcagaatccc tgcttcgtcc     7260
atttgacagg cacattatgc atgccgcttc gccttcgcgc gcgaattgat ctgctgcctc     7320
gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca     7380
gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt     7440
ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc     7500
ttaactatgc ggcatcagag cagattgtac tgagagtgca ggggatcgcg gccgcggacc     7560
ggatcctcta gagcggccgc gatcctctag agtcgaccgg gaatggcgaa tgggacgcgc     7620
cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac     7680
ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg     7740
ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt     7800
tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc     7860
cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct     7920
tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga     7980
ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga     8040
attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg     8100
aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tcagatcctt     8160
tttaacccat cacatatacc tgccgttcac tattatttag tgaaatgaga tattatgata     8220
ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac tataaaaaat     8280
acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag tctgcaaatc     8340
cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc caaacgagag     8400
tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa agcaggcaag     8460
acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca tattttaggt     8520
ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg aagaagcaga     8580
ccgctaacac agtacataaa aaaggagaca tgaacgatga acatcaaaaa gtttgcaaaa     8640
caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt     8700
gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc     8760
catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt tcctgaattc     8820
gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg     8880
ccattacaaa acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca     8940
ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc     9000
ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa     9060
ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca     9120
tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc     9180
aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc     9240
aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta     9300
cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct     9360
cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa     9420
gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca     9480
ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta     9540
gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg     9600
aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa     9660
atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc     9720
attacgtcta acgatattta catgcttggt tatgtttcta attctttaac tggcccatac     9780
aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc     9840
tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc     9900
tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc aagcttcctg     9960
ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa    10020
ttaacagtta acaaataaaa acgcaaaaga aaatgccgat ccggtttatt gactaccgga    10080
agcagtgtga ccgtgtgctt ctcaaatgcc tcaggctgtc tatgtgtgac tgttgagctg    10140
taacaagttg tctcaggtgt tcaatttcat gttctagttg ctttgtttta ctggtttcac    10200
ctgttctatt aggtgttaca tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa    10260
cagctttaaa tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt tttacaccgt    10320
tttcatctgt gcatatggac agttttccct ttgatatcta acggtgaaca gttgttctac    10380
ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc    10440
cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag    10500
aacgaaccat tgagatcatg cttactttgc atgtcactca aaaattttgc ctcaaaactg    10560
gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttacgtagg    10620
taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc    10680
tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca    10740
gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta    10800
cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc    10860
attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt    10920
tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac    10980
ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc    11040
tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg    11100
aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct    11160
ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg    11220
tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg    11280
ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga    11340
ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct    11400
aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt    11460
ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat    11520
tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa    11580
gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc    11640
cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg    11700
ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca    11760
agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg    11820
tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg    11880
gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc    11940
cgtcacgggc ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt    12000
tgctgttcag cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg    12060
acaggtcatt cagactggct aatgcaccca gtaaggcagc ggtatcatca acaggcttac    12120
ccgtcttact gtcggatcga cgctctccct tatgcgactc ctgcat                   12166

<210>  87
<211>  410
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Y155H encoding synthetic fragment

<400>  87
ctccagtcgt ctgctcttcg agagtcgagc agacaagatg gcctttgcat cgactccgga       60
ggcattgatg gggttttcct cgaacagctg gttgtaggtc tgaacgagct gaatgaacag      120
cttgtccaca tcgctgttgt cgggattgag atcgccctcg atgaggaaat gacctcgaaa      180
cttgatcatg tgtgccagag cgagatggat aagtctgaga tccgccttgt cggtggaatc      240
gacgagtttc tttcgcaggt ggtagatggt aggatacttc tcgtggtaag caacctcgtc      300
cacaatgttg ccaaagatgg gatgacgctc gtgtttcttg tcttcctcga cgaggaagga      360
ttcctccagt cgatgaaaga acgaatcgtc caccttggcc atctcgttgg                 410

<210>  88
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Y155H synthetic fragment forward primer

<400>  88
ccaacgagat ggccaaggtg                                                   20

<210>  89
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Y155H synthetic fragment reverse primer

<400>  89
ccaacgagat ggccaaggtg                                                   20

<210>  90
<211>  11794
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF97-Y155H fragment of plasmid

<400>  90
caccttggcc atctcgttgg aaaagatctc ctgcaggtag cagattcggt tcttccgtcg       60
ggtgtaacgt cgccgagcag ttcgcttgag tctggtagct tcggcagtct cgccagaatc      120
gaacaacagg gcaccaatga ggtttttctt gatggagtgt cgatcggtgt ttccgaggac      180
cttgaatttc ttggagggca ccttgtactc gtcggtgatg acagcccagc cgacagagtt      240
ggttccaatg tccaggccga tggagtattt cttgtcgaat tcccatatgg taccagctgc      300
agatctcgag ctcggatcct tatcgtcatc gtcgtacaga tcccgaccca tttgctgtcc      360
accagtcatg ctagccatac catgatgatg atgatgatga gaacccccca tggttaattc      420
ctcctgttag cccaaaaaac gggtatggag aaacagtaga gagttgcgat aaaaagcgtc      480
aggtaggatc cgctaatctt atggataaaa atgctatggc atagcaaagt gtgacgccgt      540
gcaaataatc aatgtggact tttctgccgt gattatagac acttttgtta cgcgtttttg      600
tcatggcttt ggtcccgctt tgttacagaa tgcttttaat aagcggggtt accggtttgg      660
ttagcgagaa gagccagtaa aagacgcagt gacggcaatg tctgatgcaa tatggacaat      720
tggtttcttc tctgaatggc gggagtatga aaagtatggc tgaagcgcaa aatgatcccc      780
tgctgccggg atactcgttt aatgcccatc tggtggcggg tttaacgccg attgaggcca      840
acggttatct cgattttttt atcgaccgac cgctgggaat gaaaggttat attctcaatc      900
tcaccattcg cggtcagggg gtggtgaaaa atcagggacg agaatttgtt tgccgaccgg      960
gtgatatttt gctgttcccg ccaggagaga ttcatcacta cggtcgtcat ccggaggctc     1020
gcgaatggta tcaccagtgg gtttactttc gtccgcgcgc ctactggcat gaatggctta     1080
actggccgtc aatatttgcc aatacggggt tctttcgccc ggatgaagcg caccagccgc     1140
atttcagcga cctgtttggg caaatcatta acgccgggca aggggaaggg cgctattcgg     1200
agctgctggc gataaatctg cttgagcaat tgttactgcg gcgcatggaa gcgattaacg     1260
agtcgctcca tccaccgatg gataatcggg tacgcgaggc ttgtcagtac atcagcgatc     1320
acctggcaga cagcaatttt gatatcgcca gcgtcgcaca gcatgtttgc ttgtcgccgt     1380
cgcgtctgtc acatcttttc cgccagcagt tagggattag cgtcttaagc tggcgcgagg     1440
accaacgtat cagccaggcg aagctgcttt tgagcaccac ccggatgcct atcgccaccg     1500
tcggtcgcaa tgttggtttt gacgatcaac tctatttctc gcgggtattt aaaaaatgca     1560
ccggggccag cccgagcgag ttccgtgccg gttgtgaaga aaaagtgaat gatgtagccg     1620
tcaagttgtc ataattggta acgaatcaga caattgacgg cttgacggag tagcataggg     1680
tttgcagaat ccctgcttcg tccatttgac aggcacatta tgcatgccgc ttcgccttcg     1740
cgcgcgaatt gatctgctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca     1800
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc     1860
gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta     1920
gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt     1980
gcaggggatc gcggccgcgg accggatcct ctagagcggc cgcgatcctc tagagtcgac     2040
cggtggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac     2100
gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc     2160
ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt     2220
agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg     2280
ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac     2340
gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta     2400
ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat     2460
ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact     2520
tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg     2580
tatccgctca ccgcgatcct ttttaaccca tcacatatac ctgccgttca ctattattta     2640
gtgaaatgag atattatgat attttctgaa ttgtgattaa aaaggcaact ttatgcccat     2700
gcaacagaaa ctataaaaaa tacagagaat gaaaagaaac agatagattt tttagttctt     2760
taggcccgta gtctgcaaat ccttttatga ttttctatca aacaaaagag gaaaatagac     2820
cagttgcaat ccaaacgaga gtctaataga atgaggtcga aaagtaaatc gcgcgggttt     2880
gttactgata aagcaggcaa gacctaaaat gtgtaaaggg caaagtgtat actttggcgt     2940
caccccttac atattttagg tcttttttta ttgtgcgtaa ctaacttgcc atcttcaaac     3000
aggagggctg gaagaagcag accgctaaca cagtacataa aaaaggagac atgaacgatg     3060
aacatcaaaa agtttgcaaa acaagcaaca gtattaacct ttactaccgc actgctggca     3120
ggaggcgcaa ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa ggaaacatac     3180
ggcatttccc atattacacg ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa     3240
aaatatcaag ttcctgagtt cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc     3300
ctggacgttt gggacagctg gccattacaa aacgctgacg gcactgtcgc aaactatcac     3360
ggctaccaca tcgtctttgc attagccgga gatcctaaaa atgcggatga cacatcgatt     3420
tacatgttct atcaaaaagt cggcgaaact tctattgaca gctggaaaaa cgctggccgc     3480
gtctttaaag acagcgacaa attcgatgca aatgattcta tcctaaaaga ccaaacacaa     3540
gaatggtcag gttcagccac atttacatct gacggaaaaa tccgtttatt ctacactgat     3600
ttctccggta aacattacgg caaacaaaca ctgacaactg cacaagttaa cgtatcagca     3660
tcagacagct ctttgaacat caacggtgta gaggattata aatcaatctt tgacggtgac     3720
ggaaaaacgt atcaaaatgt acagcagttc atcgatgaag gcaactacag ctcaggcgac     3780
aaccatacgc tgagagatcc tcactacgta gaagataaag gccacaaata cttagtattt     3840
gaagcaaaca ctggaactga agatggctac caaggcgaag aatctttatt taacaaagca     3900
tactatggca aaagcacatc attcttccgt caagaaagtc aaaaacttct gcaaagcgat     3960
aaaaaacgca cggctgagtt agcaaacggc gctctcggta tgattgagct aaacgatgat     4020
tacacactga aaaaagtgat gaaaccgctg attgcatcta acacagtaac agatgaaatt     4080
gaacgcgcga acgtctttaa aatgaacggc aaatggtacc tgttcactga ctcccgcgga     4140
tcaaaaatga cgattgacgg cattacgtct aacgatattt acatgcttgg ttatgtttct     4200
aattctttaa ctggcccata caagccgctg aacaaaactg gccttgtgtt aaaaatggat     4260
cttgatccta acgatgtaac ctttacttac tcacacttcg ctgtacctca agcgaaagga     4320
aacaatgtcg tgattacaag ctatatgaca aacagaggat tctacgcaga caaacaatca     4380
acgtttgcgc caagcttcct gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac     4440
agcatccttg aacaaggaca attaacagtt aacaaataaa aacgcaaaag aaaatgccga     4500
tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgattc aggctgtcta     4560
tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct     4620
ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt     4680
cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc tctgatgtat     4740
ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac     4800
ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag     4860
ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt     4920
ttttgcgtga gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa     4980
aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt     5040
cttagtccgt tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc     5100
atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact     5160
tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg     5220
taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca     5280
tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt     5340
gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag     5400
tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg     5460
aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg     5520
gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca     5580
gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg     5640
atgttcatca tctgaacgta ttggttataa gtgaacgata ccgtccgttc tttccttgta     5700
gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc     5760
tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac     5820
atacatctca attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa     5880
tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt     5940
gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt     6000
ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata     6060
aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac     6120
aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc     6180
ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat     6240
caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc     6300
agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc     6360
cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg     6420
tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc     6480
acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg     6540
tatcatcaac aggcttaccc gtcttactgt cggggatcga cgctctccct tatgcgactc     6600
ctgcaccttt cgtcttcgaa taaatacctg tgacggaaga tcacttcgca gaataaataa     6660
atcctggtgt ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg     6720
ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg     6780
tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca     6840
ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc     6900
agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa     6960
agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc     7020
tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg     7080
atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct     7140
ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt     7200
gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct     7260
cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact     7320
tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc     7380
cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta     7440
atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt     7500
attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg     7560
gcagaaattc gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc     7620
cgcttatgtc tattgctggt ctcggtaccc tgcacccaac tgatcttcag catcttttac     7680
tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat     7740
aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat     7800
ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca     7860
aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat     7920
gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt     7980
tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga     8040
taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc     8100
cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag     8160
ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc     8220
agaccgcttc tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca     8280
aaacagccaa gcttagcggc cgcttagacc tttcgctttt tcttgggatc ggctctggag     8340
tcgccaccaa gctgagacag gtcgattcgg gtctcgtaca ggccagtgat ggactggtga     8400
atcagggtgg catcgagaac ctccttggtg gatgtgtacc gctttcggtc gatagtggta     8460
tcgaagtact tgaaagctgc aggagcaccc aggttggtaa gagtaaacag gtgaatgatg     8520
ttctccgcct gttctcgaat gggtttgtcc cgatgcttgt tgtaggcaga gagcaccttg     8580
tccaagttgg catcagccag gatgactcgc ttcgaaaact cggaaatctg ctcgataatc     8640
tcgtcgaggt aatgtttgtg ctgctcaacg aagagttgct tctgttcgtt gtcctcggga     8700
gaacccttga gcttctcgta gtgagaagcc agatagagaa agttgacgta cttcgaaggc     8760
aaggcaagct cgtttccctt ctgcagctcg ccagcggagg cgagcatacg ctttcgaccg     8820
ttctccagtt cgaacagaga gtacttgggc agcttgataa tgaggtcttt cttgacctcc     8880
ttgtaaccct tggcttccaa gaagtcgatg ggattcttct cgaagctcga tcgctccatg     8940
atggtaattc cgagcagctc cttgacggac ttgagctttt tggacttgcc cttctcgacc     9000
ttcgcaacga caagcacgga ataggcgacg gtaggagaat cgaagccacc gtatttcttg     9060
ggatcccagt ctttctttcg agcgatgagc ttgtcggagt ttcgcttggg cagaatcgac     9120
tccttggaga atccgccagt ctgaacctcg gttttcttga cgatgttgac ctgaggcatc     9180
gacagaacct ttcgcacggt tgcaaagtct cgacccttgt cccacacgat ctctccagtt     9240
tcgccgttgg tctcgataag tggtctcttt cgaatctctc cgttggccaa ggtgatctcg     9300
gtcttgaaaa agttcatgat gttggagtaa aagaagtact tggcagtagc cttgccaatc     9360
tcctgttcgg acttggcaat catctttcga acgtcgtaga ccttgtaatc gccgtaaacg     9420
aactcgcttt cgagcttggg gtatttcttg atgagcgcag tgccaacgac ggcgttgagg     9480
taagcatcgt gggcatggtg gtaattgttg atctctcgca ccttgtagaa ctgaaagtcc     9540
tttcggaaat cggagaccag tttggacttg agagtaatca ccttgacctc tcggatgagc     9600
ttgtcgttct cgtcgtactt ggtgttcatc cgagaatcga gaatctgtgc gacgtgcttt     9660
gtgatctgtc tggtctcgac gagttgacgc ttgatgaagc cagccttgtc gagctcggac     9720
agaccgcctc gctcggcctt ggtaagattg tcgaactttc gctgggtaat gagcttggcg     9780
ttgagcagct gtcgccagta gttcttcatc tttttgacca cctcttcgct gggaacgttg     9840
tccgacttgc ctctgttctt gtcggatcgt gtaaggacct tgttgtcgat agaatcgtcc     9900
ttgagaaagg attgagggac aatgtggtcc acatcgtagt cgctgagacg attgatgtcc     9960
agttcctgat ccacgtacat gtctcgacca ttctgcagat agtagagata cagcttctcg    10020
ttctgcagtt gagtgttctc gacgggatgc tccttgagaa tctgggatcc cagctccttg    10080
atgccttcct cgattcgctt catccgctct cgcgagtttt tctgaccctt ttgagttgtc    10140
tggttctctc tggccatctc gatcacaatg ttctcgggct tgtgacgtcc catgaccttc    10200
accagctcgt cgacaacctt gacagtctgg agaatgcctt tcttgatggc tggcgaacca    10260
gccaggttgg caatatgttc gtgcaagctg tcgccctgac cggacacttg tgccttctgg    10320
atgtcctcct tgaaggtaag agaatcgtcg tgaatgagct gcatgaagtt tcggttggca    10380
aagccatcgg acttgagaaa gtccagaatg gtctttccgg actgcttgtc tctgatgccg    10440
ttgatgagct ttcgcgaaag tcttccccag ccggtgtatc tacgtcgctt gagttgtttc    10500
atgaccttgt cgtcgaacag gtgagcgtat gtcttgagtc gttcctcgat catctcccga    10560
tcttcgaaca gggtaagagt gagcacgatg tcctccagaa tgtcctcgtt ttcctcgttg    10620
tcgagaaaat ccttgtcctt gataatcttg agcagatcgt gataggtgcc caaagaggcg    10680
ttgaatcggt cctcaactcc ggaaatctcg acgctgtcga aacactcgat tttcttgaag    10740
tagtcctcct tgagctgctt aacagtgacc tttcggttgg tcttgaacag gagatcgaca    10800
atggctttct tctgttcgcc agacaagaag gcaggctttc gcattccctc ggtaacgtac    10860
ttgactttgg tgagttcgtt gtagactgta aagtactcgt agagcagcga atgcttggga    10920
agaaccttct cgttgggcag attcttgtcg aagttggtca ttcgctcgat gaaggactgt    10980
gcagaggcac ccttgtccac gacttcctcg aagttccagg gagtgatggt ttcctcggac    11040
tttcgagtca tccaagcaaa tcgagagttt cctctggcaa gaggaccaac atagtagggg    11100
attcgaaagg taagaatctt ctcgatcttc tctcggttgt ccttgagaaa ggggtagaag    11160
tcttcctgac gtcgaagaat ggcgtgcagc tcaccgaggt ggatctgatg aggaatgctg    11220
ccgttgtcga aggttcgttg cttccgaagc agatcctctc gattgagctt gacaagcagt    11280
tcctcggttc cgtccatctt ctcgagaatt ggcttgatga acttgtagaa ctcttcctga    11340
gaggctccgc cgtcgatgta tccagcgtag ccgttcttcg actgatcgaa aaagatctcc    11400
ttgtacttct cgggcagttg ctgtcggaca agagccttga gcagtgtgag atcctgatgg    11460
tgctcgtcgt atcgcttgat catggaggca gaaaggggag cctttgtgat ctcggtgttg    11520
actcgcagaa tgtcagacaa gagaatagca tccgaaaggt tcttggcagc gagaaacagg    11580
tcggcgtact gatcgccaat ctgtgcaagc aggttgtcga ggtcatcgtc gtaggtgtcc    11640
ttggacagct ggagcttggc gtcctccgcc agatcgaagt tggacttgaa gttgggtgtg    11700
agaccaagag aaagggcaat gaggttgcca aacagtccgt tctttttctc gccaggaagt    11760
tgggcaatga ggttctccag tcgtctgctc ttcg                                11794

<210>  91
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF97-Y155H fragment forward primer

<400>  91
caccttggcc atctcgttgg                                                   20

<210>  92
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF97-Y155H fragment reverse primer

<400>  92
cgaagagcag acgactggag                                                   20

<210>  93
<211>  12164
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF861

<400>  93
cctttcgtct tcgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct       60
ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat      120
cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt      180
tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga      240
tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca      300
gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc      360
gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg      420
aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt      480
gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt      540
gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac      600
ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc      660
aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc      720
gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg      780
gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa      840
ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg      900
tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtctcgg taccctgcac ccaactgatc ttcagcatct tttactttca     1080
ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg     1140
cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc     1200
agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaaaga     1260
gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat ttgatgcctg     1320
gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc aacgttcaaa     1380
tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa     1440
cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac     1500
tctcgcatgg ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg     1560
catggggtca ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc     1620
gcttctgcgt tctgatttaa tctgtatcag gctgaaaatc ttctctcatc cgccaaaaca     1680
gccaagctta gcggccgctt agacctttcg ctttttcttg ggatcggctc tggagtcgcc     1740
accaagctga gacaggtcga ttcgggtctc gtacaggcca gtgatggact ggtgaatcag     1800
ggtggcatcg agaacctcct tggtggatgt gtaccgcttt cggtcgatag tggtatcgaa     1860
gtacttgaaa gctgcaggag cacccaggtt ggtaagagta aacaggtgaa tgatgttctc     1920
cgcctgttct cgaatgggtt tgtcccgatg cttgttgtag gcagagagca ccttgtccaa     1980
gttggcatca gccaggatga ctcgcttcga aaactcggaa atctgctcga taatctcgtc     2040
gaggtaatgt ttgtgctgct caacgaagag ttgcttctgt tcgttgtcct cgggagaacc     2100
cttgagcttc tcgtagtgag aagccagata gagaaagttg acgtacttcg aaggcaaggc     2160
aagctcgttt cccttctgca gctcgccagc ggaggcgagc atacgctttc gaccgttctc     2220
cagttcgaac agagagtact tgggcagctt gataatgagg tctttcttga cctccttgta     2280
acccttggct tccaagaagt cgatgggatt cttctcgaag ctcgatcgct ccatgatggt     2340
aattccgagc agctccttga cggacttgag ctttttggac ttgcccttct cgaccttcgc     2400
aacgacaagc acggaatagg cgacggtagg agaatcgaag ccaccgtatt tcttgggatc     2460
ccagtctttc tttcgagcga tgagcttgtc ggagtttcgc ttgggcagaa tcgactcctt     2520
ggagaatccg ccagtctgaa cctcggtttt cttgacgatg ttgacctgag gcatcgacag     2580
aacctttcgc acggttgcaa agtctcgacc cttgtcccac acgatctctc cagtttcgcc     2640
gttggtctcg ataagtggtc tctttcgaat ctctccgttg gccaaggtga tctcggtctt     2700
gaaaaagttc atgatgttgg agtaaaagaa gtacttggca gtagccttgc caatctcctg     2760
ttcggacttg gcaatcatct ttcgaacgtc gtagaccttg taatcgccgt aaacgaactc     2820
gctttcgagc ttggggtatt tcttgatgag cgcagtgcca acgacggcgt tgaggtaagc     2880
atcgtgggca tggtggtaat tgttgatctc tcgcaccttg tagaactgaa agtcctttcg     2940
gaaatcggag accagtttgg acttgagagt aatcaccttg acctctcgga tgagcttgtc     3000
gttctcgtcg tacttggtgt tcatccgaga atcgagaatc tgtgcgacgt gctttgtgat     3060
ctgtctggtc tcgacgagtt gacgcttgat gaagccagcc ttgtcgagct cggacagacc     3120
gcctcgctcg gccttggtaa gattgtcgaa ctttcgctgg gtaatgagct tggcgttgag     3180
cagctgtcgc cagtagttct tcatcttttt gaccacctct tcgctgggaa cgttgtccga     3240
cttgcctctg ttcttgtcgg atcgtgtaag gaccttgttg tcgatagaat cgtccttgag     3300
aaaggattga gggacaatgt ggtccacatc gtagtcgctg agacgattga tgtccagttc     3360
ctgatccacg tacatgtctc gaccattctg cagatagtag agatacagct tctcgttctg     3420
cagttgagtg ttctcgacgg gatgctcctt gagaatctgg gatcccagct ccttgatgcc     3480
ttcctcgatt cgcttcatcc gctctcgcga gtttttctga cccttttgag ttgtctggtt     3540
ctctctggcc atctcgatca caatgttctc gggcttgtga cgtcccatga ccttcaccag     3600
ctcgtcgaca accttgacag tctggagaat gcctttcttg atggctggcg aaccagccag     3660
gttggcaata tgttcgtgca agctgtcgcc ctgaccggac acttgtgcct tctggatgtc     3720
ctccttgaag gtaagagaat cgtcgtgaat gagctgcatg aagtttcggt tggcaaagcc     3780
atcggacttg agaaagtcca gaatggtctt tccggactgc ttgtctctga tgccgttgat     3840
gagctttcgc gaaagtcttc cccagccggt gtatctacgt cgcttgagtt gtttcatgac     3900
cttgtcgtcg aacaggtgag cgtatgtctt gagtcgttcc tcgatcatct cccgatcttc     3960
gaacagggta agagtgagca cgatgtcctc cagaatgtcc tcgttttcct cgttgtcgag     4020
aaaatccttg tccttgataa tcttgagcag atcgtgatag gtgcccaaag aggcgttgaa     4080
tcggtcctca actccggaaa tctcgacgct gtcgaaacac tcgattttct tgaagtagtc     4140
ctccttgagc tgcttaacag tgacctttcg gttggtcttg aacaggagat cgacaatggc     4200
tttcttctgt tcgccagaca agaaggcagg ctttcgcatt ccctcggtaa cgtacttgac     4260
tttggtgagt tcgttgtaga ctgtaaagta ctcgtagagc agcgaatgct tgggaagaac     4320
cttctcgttg ggcagattct tgtcgaagtt ggtcattcgc tcgatgaagg actgtgcaga     4380
ggcacccttg tccacgactt cctcgaagtt ccagggagtg atggtttcct cggactttcg     4440
agtcatccaa gcaaatcgag agtttcctct ggcaagagga ccaacatagt aggggattcg     4500
aaaggtaaga atcttctcga tcttctctcg gttgtccttg agaaaggggt agaagtcttc     4560
ctgacgtcga agaatggcgt gcagctcacc gaggtggatc tgatgaggaa tgctgccgtt     4620
gtcgaaggtt cgttgcttcc gaagcagatc ctctcgattg agcttgacaa gcagttcctc     4680
ggttccgtcc atcttctcga gaattggctt gatgaacttg tagaactctt cctgagaggc     4740
tccgccgtcg atgtatccag cgtagccgtt cttcgactga tcgaaaaaga tctccttgta     4800
cttctcgggc agttgctgtc ggacaagagc cttgagcagt gtgagatcct gatggtgctc     4860
gtcgtatcgc ttgatcatgg aggcagaaag gggagccttt gtgatctcgg tgttgactcg     4920
cagaatgtca gacaagagaa tagcatccga aaggttcttg gcagcgagaa acaggtcggc     4980
gtactgatcg ccaatctgtg caagcaggtt gtcgaggtca tcgtcgtagg tgtccttgga     5040
cagctggagc ttggcgtcct ccgccagatc gaagttggac ttgaagttgg gtgtgagacc     5100
aagagaaagg gcaatgaggt tgccaaacag tccgttcttt ttctcgccag gaagttgggc     5160
aatgaggttc tccagtcgtc tgctcttcga gagtcgagca gacaagatgg cctttgcatc     5220
gactccggag gcattgatgg ggttttcctc gaacagctgg ttgtaggtct gaacgagctg     5280
aatgaacagc ttgtccacat cgctgttgtc gggattgaga tcgccctcga tgaggaaatg     5340
acctcgaaac ttgatcatgt gtgccagagc gagatggata agtctgagat ccgccttgtc     5400
ggtggaatcg acgagtttct ttcgcaggtg gtagatggta ggatacttct cgtggtaagc     5460
aacctcgtcc acaatgttgc caaagatggg atgacgctcg tgtttcttgt cttcctcgac     5520
gaggaaggat tcctccagtc gatgaaagaa cgaatcgtcc accttggcca tctcgttgga     5580
aaagatctcc tgcaggtagc agattcggtt cttccgtcgg gtgtaacgtc gccgagcagt     5640
tcgcttgagt ctggtagctt cggcagtctc gccagaatcg aacaacaggg caccaatgag     5700
gtttttcttg atggagtgtc gatcggtgtt tccgaggacc ttgaatttct tggagggcac     5760
cttgtactcg tcggtgatga cagcccagcc gacagagttg gttccaatgt ccaggccgat     5820
ggagtatttc ttgtcgaatt cccatatggt accagctgca gatctcgagc tcggatcctt     5880
atcgtcatcg tcgtacagat cccgacccat ttgctgtcca ccagtcatgc tagccatacc     5940
atgatgatga tgatgatgag aaccccccat ggttaattcc tcctgttagc ccaaaaaacg     6000
ggtatggaga aacagtagag agttgcgata aaaagcgtca ggtaggatcc gctaatctta     6060
tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca atgtggactt     6120
ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg gtcccgcttt     6180
gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag agccagtaaa     6240
agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttcttct ctgaatggcg     6300
ggagtatgaa aagtatggct gaagcgcaaa atgatcccct gctgccggga tactcgttta     6360
atgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc gattttttta     6420
tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc ggtcaggggg     6480
tggtgaaaaa tcagggacga gaatttgttt gccgaccggg tgatattttg ctgttcccgc     6540
caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat caccagtggg     6600
tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca atatttgcca     6660
atacggggtt ctttcgcccg gatgaagcgc accagccgca tttcagcgac ctgtttgggc     6720
aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg ataaatctgc     6780
ttgagcaatt gttactgcgg cgcatggaag cgattaacga gtcgctccat ccaccgatgg     6840
ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac agcaattttg     6900
atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca catcttttcc     6960
gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc agccaggcga     7020
agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat gttggttttg     7080
acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc ccgagcgagt     7140
tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca taattggtaa     7200
cgaatcagac aattgacggc ttgacggagt agcatagggt ttgcagaatc cctgcttcgt     7260
ccatttgaca ggcacattat gcatgccgct tcgccttcgc gcgcgaattg atctgctgcc     7320
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca     7380
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg     7440
ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg     7500
gcttaactat gcggcatcag agcagattgt actgagagtg caggggatcg cggccgcgga     7560
ccggatcctc tagagcggcc gcgatcctct agagtcgacc ggtggcgaat gggacgcgcc     7620
ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact     7680
tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc     7740
cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt     7800
acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc     7860
ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt     7920
gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat     7980
tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa     8040
ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga     8100
acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcac cgcgatcctt     8160
tttaacccat cacatatacc tgccgttcac tattatttag tgaaatgaga tattatgata     8220
ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac tataaaaaat     8280
acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag tctgcaaatc     8340
cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc caaacgagag     8400
tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa agcaggcaag     8460
acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca tattttaggt     8520
ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg aagaagcaga     8580
ccgctaacac agtacataaa aaaggagaca tgaacgatga acatcaaaaa gtttgcaaaa     8640
caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt     8700
gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc     8760
catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt tcctgagttc     8820
gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg     8880
ccattacaaa acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca     8940
ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc     9000
ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa     9060
ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca     9120
tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc     9180
aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc     9240
aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta     9300
cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct     9360
cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa     9420
gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca     9480
ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta     9540
gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg     9600
aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa     9660
atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc     9720
attacgtcta acgatattta catgcttggt tatgtttcta attctttaac tggcccatac     9780
aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc     9840
tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc     9900
tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc aagcttcctg     9960
ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa    10020
ttaacagtta acaaataaaa acgcaaaaga aaatgccgat attgactacc ggaagcagtg    10080
tgaccgtgtg cttctcaaat gcctgattca ggctgtctat gtgtgactgt tgagctgtaa    10140
caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg    10200
ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag    10260
ctttaaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt    10320
catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt    10380
tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt    10440
ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac    10500
gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg    10560
agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag    10620
gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca    10680
agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc    10740
gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt    10800
attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt    10860
aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt    10920
gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta    10980
acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct    11040
tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa    11100
atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg    11160
gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgaacgtat    11220
tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg    11280
agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta    11340
atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg    11400
tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc    11460
ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct    11520
gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg    11580
gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct    11640
gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg    11700
tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc    11760
tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct    11820
ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca    11880
ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt    11940
cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc    12000
tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca    12060
ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg    12120
tcttactgtc ggggatcgac gctctccctt atgcgactcc tgca                     12164

<210>  94
<211>  918
<212>  DNA
<213>  E. coli

<400>  94
atgaacttca gacgcctgaa atacttcgta aaaattgtag atattggtag cctgacccag       60
gctgctgaag tattgcatat cgcacaacca gcgctcagcc agcaggttgc cacactggaa      120
ggtgagttaa atcaacaact tttgatccgt acaaagcggg gcgttacacc aacagacgcc      180
ggaaaaattc tctataccca tgcgcgggcc attttacgtc agtgtgaaca ggcccaactg      240
gcggtgcata acgttggtca ggcattatcg gggcaagtct cgattggctt tgcaccagga      300
accgctgcgt catccatcac catgccctta ttacaggcgg ttcgcgctga atttccggag      360
atcgttatct atcttcatga aaatagtggt gcagtgctta acgaaaaatt gataaatcac      420
caactcgata tggcggtgat ttatgagcat tcccctgtgg ctggtgtatc cagtcaggct      480
ttgctgaaag aagatctttt tctggtagga actcaagatt gcccggggca aagcgttgat      540
gtgaatgcta ttgcgcaaat gaacctcttt ctccccagtg attacagtgc tattagactt      600
cgtgttgatg aggctttttc cctacggcga ctcacggcaa aagttattgg tgaaattgag      660
tctattgcca cgcttaccgc agcgattgcc agcggcatgg gcgttgcagt attacccgaa      720
tcggccgcgc gttcgttatg tggcgcagta aatgggtgga tgtcacgcat taccactcct      780
tccatgagtc tctctttgtc attaaattta cccgccagag cgaacttatc gccacaggca      840
caggcagtga aagagttgtt aatgtcagtg attagttctc cagtgatgga aaaaaggcag      900
tggcaattgg tgagctaa                                                    918

<210>  95
<211>  20
<212>  DNA
<213>  E. coli

<400>  95
tatgcaatac ttcagcagcc                                                   20

<210>  96
<211>  23
<212>  DNA
<213>  E. coli

<400>  96
tatgcaatac ttcagcagcc tgg                                               23

<210>  97
<211>  20
<212>  DNA
<213>  E. coli

<400>  97
acaaccagcg ctcagccagc                                                   20

<210>  98
<211>  23
<212>  DNA
<213>  E. coli

<400>  98
acaaccagcg ctcagccagc agg                                               23

<210>  99
<211>  56
<212>  DNA
<213>  N25 phage

<400>  99
aagaatcata aaaaatttat ttgctttcag gaaaattttt ctgtataata gattca           56

<210>  100
<211>  247
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized nac target site 1 gRNA expression cassette

<400>  100
aagaatcata aaaaatttat ttgctttcag gaaaattttt ctgtataata gattcatatg       60
caatacttca gcagccgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt      120
atcaacttga aaaagtggca ccgagtcggt gcgactcctg ttgatagatc cagtaatgac      180
ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg      240
tgagaat                                                                247

<210>  101
<211>  247
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized nac target site 2 gRNA expression cassette

<400>  101
aagaatcata aaaaatttat ttgctttcag gaaaattttt ctgtataata gattcaacaa       60
ccagcgctca gccagcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt      120
atcaacttga aaaagtggca ccgagtcggt gcgactcctg ttgatagatc cagtaatgac      180
ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg      240
tgagaat                                                                247

<210>  102
<211>  500
<212>  DNA
<213>  E. coli

<400>  102
gaagttcatg ttgcctccgg tttttaagaa tcggcccaag tgccgccatt acttacaacc       60
agattgcaag atgcttgcca gttttatttt ggtgttgatg tacaagctaa ccaactgtca      120
aataagagat tatgatagat tcgtcatttg ctcctttaat cagctgtcgc gttcccctgc      180
cctataaaag gagggtatgc accacgatgg ttcattaccc aataagattg aaagctcacc      240
actttgttga aattgacagc aaacaaacaa aaaaatgcat ttcacccttt gacatcacca      300
tgcactgcca ttaatatgcg ccccgttcac acgattcctc tgtagttcag tcggtagaac      360
ggcggactgt taatccgtat gtcactggtt cgagtccagt cagaggagcc aaattcaaaa      420
aagcctgctt tctagcaggc tttttgcttt ctaattacca acgctcttaa aacatctgtc      480
ttgaaccaga actaatttgc                                                  500

<210>  103
<211>  500
<212>  DNA
<213>  E. coli

<400>  103
cgtcccctgg attagctcga gccgaacctc cgggaaaagt tcgcgaaaag ctttaatgac       60
ctctggcaag ctataacgtg cctgagtatg cgtcgttgca atagtgagaa cgccagacgt      120
atcgttggta aacaggtctg caagccgacg aacattactg gcttcattca gaatacgttc      180
tgcaatgacc agtaatgctt tgcccggttc agtcatgccc agcagtcgct tacctcgtcg      240
aacaaatatt tcgatgccaa gttcatcctc cagttcccga atatgacggc tgacgcctga      300
ctgtgaggta aaaagcatat tcgcaacctc tgtcaggttg taatcctgac gtgcagcctc      360
gcggattatc tttagttgtt ggaaattcac ggtaaactcc gggcagttca gatttcccgt      420
tattgttaaa gtctaatgcc cggcataaca aataataaaa acccgcatct tattccatcc      480
cgatataaca cttagctcac                                                  500

<210>  104
<211>  1000
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized nac deletion editing template

<400>  104
cgtcccctgg attagctcga gccgaacctc cgggaaaagt tcgcgaaaag ctttaatgac       60
ctctggcaag ctataacgtg cctgagtatg cgtcgttgca atagtgagaa cgccagacgt      120
atcgttggta aacaggtctg caagccgacg aacattactg gcttcattca gaatacgttc      180
tgcaatgacc agtaatgctt tgcccggttc agtcatgccc agcagtcgct tacctcgtcg      240
aacaaatatt tcgatgccaa gttcatcctc cagttcccga atatgacggc tgacgcctga      300
ctgtgaggta aaaagcatat tcgcaacctc tgtcaggttg taatcctgac gtgcagcctc      360
gcggattatc tttagttgtt ggaaattcac ggtaaactcc gggcagttca gatttcccgt      420
tattgttaaa gtctaatgcc cggcataaca aataataaaa acccgcatct tattccatcc      480
cgatataaca cttagctcac gaagttcatg ttgcctccgg tttttaagaa tcggcccaag      540
tgccgccatt acttacaacc agattgcaag atgcttgcca gttttatttt ggtgttgatg      600
tacaagctaa ccaactgtca aataagagat tatgatagat tcgtcatttg ctcctttaat      660
cagctgtcgc gttcccctgc cctataaaag gagggtatgc accacgatgg ttcattaccc      720
aataagattg aaagctcacc actttgttga aattgacagc aaacaaacaa aaaaatgcat      780
ttcacccttt gacatcacca tgcactgcca ttaatatgcg ccccgttcac acgattcctc      840
tgtagttcag tcggtagaac ggcggactgt taatccgtat gtcactggtt cgagtccagt      900
cagaggagcc aaattcaaaa aagcctgctt tctagcaggc tttttgcttt ctaattacca      960
acgctcttaa aacatctgtc ttgaaccaga actaatttgc                           1000

<210>  105
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  5' pRF97 or pRF861 identity fragment

<400>  105
cgaatcagac aattgacggc                                                   20

<210>  106
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  3' pRF97 or pRF861 identity fragment

<400>  106
gcttccggta gtcaataaac c                                                 21

<210>  107
<211>  1288
<212>  DNA
<213>  artificial sequence

<220>
<223>  nacETsite1

<400>  107
cgaatcagac aattgacggc cgtcccctgg attagctcga gccgaacctc cgggaaaagt       60
tcgcgaaaag ctttaatgac ctctggcaag ctataacgtg cctgagtatg cgtcgttgca      120
atagtgagaa cgccagacgt atcgttggta aacaggtctg caagccgacg aacattactg      180
gcttcattca gaatacgttc tgcaatgacc agtaatgctt tgcccggttc agtcatgccc      240
agcagtcgct tacctcgtcg aacaaatatt tcgatgccaa gttcatcctc cagttcccga      300
atatgacggc tgacgcctga ctgtgaggta aaaagcatat tcgcaacctc tgtcaggttg      360
taatcctgac gtgcagcctc gcggattatc tttagttgtt ggaaattcac ggtaaactcc      420
gggcagttca gatttcccgt tattgttaaa gtctaatgcc cggcataaca aataataaaa      480
acccgcatct tattccatcc cgatataaca cttagctcac gaagttcatg ttgcctccgg      540
tttttaagaa tcggcccaag tgccgccatt acttacaacc agattgcaag atgcttgcca      600
gttttatttt ggtgttgatg tacaagctaa ccaactgtca aataagagat tatgatagat      660
tcgtcatttg ctcctttaat cagctgtcgc gttcccctgc cctataaaag gagggtatgc      720
accacgatgg ttcattaccc aataagattg aaagctcacc actttgttga aattgacagc      780
aaacaaacaa aaaaatgcat ttcacccttt gacatcacca tgcactgcca ttaatatgcg      840
ccccgttcac acgattcctc tgtagttcag tcggtagaac ggcggactgt taatccgtat      900
gtcactggtt cgagtccagt cagaggagcc aaattcaaaa aagcctgctt tctagcaggc      960
tttttgcttt ctaattacca acgctcttaa aacatctgtc ttgaaccaga actaatttgc     1020
aagaatcata aaaaatttat ttgctttcag gaaaattttt ctgtataata gattcatatg     1080
caatacttca gcagccgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1140
atcaacttga aaaagtggca ccgagtcggt gcgactcctg ttgatagatc cagtaatgac     1200
ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg     1260
tgagaatggt ttattgacta ccggaagc                                        1288

<210>  108
<211>  1288
<212>  DNA
<213>  artificial sequence

<220>
<223>  nacETsite2

<400>  108
cgaatcagac aattgacggc cgtcccctgg attagctcga gccgaacctc cgggaaaagt       60
tcgcgaaaag ctttaatgac ctctggcaag ctataacgtg cctgagtatg cgtcgttgca      120
atagtgagaa cgccagacgt atcgttggta aacaggtctg caagccgacg aacattactg      180
gcttcattca gaatacgttc tgcaatgacc agtaatgctt tgcccggttc agtcatgccc      240
agcagtcgct tacctcgtcg aacaaatatt tcgatgccaa gttcatcctc cagttcccga      300
atatgacggc tgacgcctga ctgtgaggta aaaagcatat tcgcaacctc tgtcaggttg      360
taatcctgac gtgcagcctc gcggattatc tttagttgtt ggaaattcac ggtaaactcc      420
gggcagttca gatttcccgt tattgttaaa gtctaatgcc cggcataaca aataataaaa      480
acccgcatct tattccatcc cgatataaca cttagctcac gaagttcatg ttgcctccgg      540
tttttaagaa tcggcccaag tgccgccatt acttacaacc agattgcaag atgcttgcca      600
gttttatttt ggtgttgatg tacaagctaa ccaactgtca aataagagat tatgatagat      660
tcgtcatttg ctcctttaat cagctgtcgc gttcccctgc cctataaaag gagggtatgc      720
accacgatgg ttcattaccc aataagattg aaagctcacc actttgttga aattgacagc      780
aaacaaacaa aaaaatgcat ttcacccttt gacatcacca tgcactgcca ttaatatgcg      840
ccccgttcac acgattcctc tgtagttcag tcggtagaac ggcggactgt taatccgtat      900
gtcactggtt cgagtccagt cagaggagcc aaattcaaaa aagcctgctt tctagcaggc      960
tttttgcttt ctaattacca acgctcttaa aacatctgtc ttgaaccaga actaatttgc     1020
aagaatcata aaaaatttat ttgctttcag gaaaattttt ctgtataata gattcaacaa     1080
ccagcgctca gccagcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1140
atcaacttga aaaagtggca ccgagtcggt gcgactcctg ttgatagatc cagtaatgac     1200
ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg     1260
tgagaatggt ttattgacta ccggaagc                                        1288

<210>  109
<211>  9322
<212>  DNA
<213>  Artificial sequence

<220>
<223>   pRF97-cassette

<400>  109
ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctc aggctgtcta       60
tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct      120
ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt      180
cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc tctgatgtat      240
ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatctaac      300
ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag      360
ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt      420
ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat gtcactcaaa      480
aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt      540
cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc      600
atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact      660
tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg      720
taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca      780
tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt      840
gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag      900
tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg      960
aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg     1020
gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca     1080
gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg     1140
atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta     1200
gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc     1260
tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac     1320
atacatctca attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa     1380
tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt     1440
gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt     1500
ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata     1560
aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac     1620
aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc     1680
ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat     1740
caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc     1800
agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc     1860
cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg     1920
tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc     1980
acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg     2040
tatcatcaac aggcttaccc gtcttactgt cggatcgacg ctctccctta tgcgactcct     2100
gcatcccttt cgtcttcgaa taaatacctg tgacggaaga tcacttcgca gaataaataa     2160
atcctggtgt ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg     2220
ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg     2280
tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca     2340
ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc     2400
agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa     2460
agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc     2520
tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg     2580
atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct     2640
ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt     2700
gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct     2760
cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact     2820
tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc     2880
cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta     2940
atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt     3000
attggtgccc ttaaacgcct ggtgctacgc ctgaataagt gataataagc ggatgaatgg     3060
cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc     3120
gcttatgtct attgctggtt tatcggtacc ccccaactga tcttcagcat cttttacttt     3180
caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag     3240
ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta     3300
tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaaa     3360
gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc     3420
tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca     3480
aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa     3540
aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct     3600
actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc     3660
ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga     3720
ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa     3780
cagccaagct tagcggccgc ttagaccttt cgctttttct tgggatcggc tctggagtcg     3840
ccaccaagct gagacaggtc gattcgggtc tcgtacaggc cagtgatgga ctggtgaatc     3900
agggtggcat cgagaacctc cttggtggat gtgtaccgct ttcggtcgat agtggtatcg     3960
aagtacttga aagctgcagg agcacccagg ttggtaagag taaacaggtg aatgatgttc     4020
tccgcctgtt ctcgaatggg tttgtcccga tgcttgttgt aggcagagag caccttgtcc     4080
aagttggcat cagccaggat gactcgcttc gaaaactcgg aaatctgctc gataatctcg     4140
tcgaggtaat gtttgtgctg ctcaacgaag agttgcttct gttcgttgtc ctcgggagaa     4200
cccttgagct tctcgtagtg agaagccaga tagagaaagt tgacgtactt cgaaggcaag     4260
gcaagctcgt ttcccttctg cagctcgcca gcggaggcga gcatacgctt tcgaccgttc     4320
tccagttcga acagagagta cttgggcagc ttgataatga ggtctttctt gacctccttg     4380
taacccttgg cttccaagaa gtcgatggga ttcttctcga agctcgatcg ctccatgatg     4440
gtaattccga gcagctcctt gacggacttg agctttttgg acttgccctt ctcgaccttc     4500
gcaacgacaa gcacggaata ggcgacggta ggagaatcga agccaccgta tttcttggga     4560
tcccagtctt tctttcgagc gatgagcttg tcggagtttc gcttgggcag aatcgactcc     4620
ttggagaatc cgccagtctg aacctcggtt ttcttgacga tgttgacctg aggcatcgac     4680
agaacctttc gcacggttgc aaagtctcga cccttgtccc acacgatctc tccagtttcg     4740
ccgttggtct cgataagtgg tctctttcga atctctccgt tggccaaggt gatctcggtc     4800
ttgaaaaagt tcatgatgtt ggagtaaaag aagtacttgg cagtagcctt gccaatctcc     4860
tgttcggact tggcaatcat ctttcgaacg tcgtagacct tgtaatcgcc gtaaacgaac     4920
tcgctttcga gcttggggta tttcttgatg agcgcagtgc caacgacggc gttgaggtaa     4980
gcatcgtggg catggtggta attgttgatc tctcgcacct tgtagaactg aaagtccttt     5040
cggaaatcgg agaccagttt ggacttgaga gtaatcacct tgacctctcg gatgagcttg     5100
tcgttctcgt cgtacttggt gttcatccga gaatcgagaa tctgtgcgac gtgctttgtg     5160
atctgtctgg tctcgacgag ttgacgcttg atgaagccag ccttgtcgag ctcggacaga     5220
ccgcctcgct cggccttggt aagattgtcg aactttcgct gggtaatgag cttggcgttg     5280
agcagctgtc gccagtagtt cttcatcttt ttgaccacct cttcgctggg aacgttgtcc     5340
gacttgcctc tgttcttgtc ggatcgtgta aggaccttgt tgtcgataga atcgtccttg     5400
agaaaggatt gagggacaat gtggtccaca tcgtagtcgc tgagacgatt gatgtccagt     5460
tcctgatcca cgtacatgtc tcgaccattc tgcagatagt agagatacag cttctcgttc     5520
tgcagttgag tgttctcgac gggatgctcc ttgagaatct gggatcccag ctccttgatg     5580
ccttcctcga ttcgcttcat ccgctctcgc gagtttttct gacccttttg agttgtctgg     5640
ttctctctgg ccatctcgat cacaatgttc tcgggcttgt gacgtcccat gaccttcacc     5700
agctcgtcga caaccttgac agtctggaga atgcctttct tgatggctgg cgaaccagcc     5760
aggttggcaa tatgttcgtg caagctgtcg ccctgaccgg acacttgtgc cttctggatg     5820
tcctccttga aggtaagaga atcgtcgtga atgagctgca tgaagtttcg gttggcaaag     5880
ccatcggact tgagaaagtc cagaatggtc tttccggact gcttgtctct gatgccgttg     5940
atgagctttc gcgaaagtct tccccagccg gtgtatctac gtcgcttgag ttgtttcatg     6000
accttgtcgt cgaacaggtg agcgtatgtc ttgagtcgtt cctcgatcat ctcccgatct     6060
tcgaacaggg taagagtgag cacgatgtcc tccagaatgt cctcgttttc ctcgttgtcg     6120
agaaaatcct tgtccttgat aatcttgagc agatcgtgat aggtgcccaa agaggcgttg     6180
aatcggtcct caactccgga aatctcgacg ctgtcgaaac actcgatttt cttgaagtag     6240
tcctccttga gctgcttaac agtgaccttt cggttggtct tgaacaggag atcgacaatg     6300
gctttcttct gttcgccaga caagaaggca ggctttcgca ttccctcggt aacgtacttg     6360
actttggtga gttcgttgta gactgtaaag tactcgtaga gcagcgaatg cttgggaaga     6420
accttctcgt tgggcagatt cttgtcgaag ttggtcattc gctcgatgaa ggactgtgca     6480
gaggcaccct tgtccacgac ttcctcgaag ttccagggag tgatggtttc ctcggacttt     6540
cgagtcatcc aagcaaatcg agagtttcct ctggcaagag gaccaacata gtaggggatt     6600
cgaaaggtaa gaatcttctc gatcttctct cggttgtcct tgagaaaggg gtagaagtct     6660
tcctgacgtc gaagaatggc gtgcagctca ccgaggtgga tctgatgagg aatgctgccg     6720
ttgtcgaagg ttcgttgctt ccgaagcaga tcctctcgat tgagcttgac aagcagttcc     6780
tcggttccgt ccatcttctc gagaattggc ttgatgaact tgtagaactc ttcctgagag     6840
gctccgccgt cgatgtatcc agcgtagccg ttcttcgact gatcgaaaaa gatctccttg     6900
tacttctcgg gcagttgctg tcggacaaga gccttgagca gtgtgagatc ctgatggtgc     6960
tcgtcgtatc gcttgatcat ggaggcagaa aggggagcct ttgtgatctc ggtgttgact     7020
cgcagaatgt cagacaagag aatagcatcc gaaaggttct tggcagcgag aaacaggtcg     7080
gcgtactgat cgccaatctg tgcaagcagg ttgtcgaggt catcgtcgta ggtgtccttg     7140
gacagctgga gcttggcgtc ctccgccaga tcgaagttgg acttgaagtt gggtgtgaga     7200
ccaagagaaa gggcaatgag gttgccaaac agtccgttct ttttctcgcc aggaagttgg     7260
gcaatgaggt tctccagtcg tctgctcttc gagagtcgag cagacaagat ggcctttgca     7320
tcgactccgg aggcattgat ggggttttcc tcgaacagct ggttgtaggt ctgaacgagc     7380
tgaatgaaca gcttgtccac atcgctgttg tcgggattga gatcgccctc gatgaggaaa     7440
tgacctcgaa acttgatcat gtgtgccaga gcgaggtaga taagtctgag atccgccttg     7500
tcggtggaat cgacgagttt ctttcgcagg tggtagatgg taggatactt ctcgtggtaa     7560
gcaacctcgt ccacaatgtt gccaaagatg ggatgacgct cgtgtttctt gtcttcctcg     7620
acgaggaagg attcctccag tcgatgaaag aacgaatcgt ccaccttggc catctcgttg     7680
gaaaagatct cctgcaggta gcagattcgg ttcttccgtc gggtgtaacg tcgccgagca     7740
gttcgcttga gtctggtagc ttcggcagtc tcgccagaat cgaacaacag ggcaccaatg     7800
aggtttttct tgatggagtg tcgatcggtg tttccgagga ccttgaattt cttggagggc     7860
accttgtact cgtcggtgat gacagcccag ccgacagagt tggttccaat gtccaggccg     7920
atggagtatt tcttgtcgaa ttcccatatg gtaccagctg cagatctcga gctcggatcc     7980
ttatcgtcat cgtcgtacag atcccgaccc atttgctgtc caccagtcat gctagccata     8040
ccatgatgat gatgatgatg agaacccccc atggttaatt cctcctgtta gcccaaaaaa     8100
cgggtatgga gaaacagtag agagttgcga taaaaagcgt caggtaggat ccgctaatct     8160
tatggataaa aatgctatgg catagcaaag tgtgacgccg tgcaaataat caatgtggac     8220
ttttctgccg tgattataga cacttttgtt acgcgttttt gtcatggctt tggtcccgct     8280
ttgttacaga atgcttttaa taagcggggt taccggtttg gttagcgaga agagccagta     8340
aaagacgcag tgacggcaat gtctgatgca atatggacaa ttggtttctt ctctgaatgg     8400
cgggagtatg aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt     8460
taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc tcgatttttt     8520
tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc gcggtcaggg     8580
ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt tgctgttccc     8640
gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt atcaccagtg     8700
ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt caatatttgc     8760
caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg acctgtttgg     8820
gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg cgataaatct     8880
gcttgagcaa ttgttactgc ggcgcatgga agcgattaac gagtcgctcc atccaccgat     8940
ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag acagcaattt     9000
tgatatcgcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt     9060
ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc     9120
gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca atgttggttt     9180
tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca gcccgagcga     9240
gttccgtgcc ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt cataattggt     9300
aacgaatcag acaattgacg gc                                              9322

<210>  110
<211>  9322
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF861-cassette

<400>  110
ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctc aggctgtcta       60
tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct      120
ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt      180
cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc tctgatgtat      240
ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatctaac      300
ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag      360
ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt      420
ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat gtcactcaaa      480
aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt      540
cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc      600
atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact      660
tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg      720
taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca      780
tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt      840
gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag      900
tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg      960
aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg     1020
gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca     1080
gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg     1140
atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta     1200
gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc     1260
tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac     1320
atacatctca attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa     1380
tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt     1440
gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt     1500
ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata     1560
aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac     1620
aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc     1680
ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat     1740
caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc     1800
agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc     1860
cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg     1920
tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc     1980
acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg     2040
tatcatcaac aggcttaccc gtcttactgt cggatcgacg ctctccctta tgcgactcct     2100
gcatcccttt cgtcttcgaa taaatacctg tgacggaaga tcacttcgca gaataaataa     2160
atcctggtgt ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg     2220
ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg     2280
tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca     2340
ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc     2400
agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa     2460
agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc     2520
tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg     2580
atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct     2640
ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt     2700
gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct     2760
cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact     2820
tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc     2880
cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta     2940
atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt     3000
attggtgccc ttaaacgcct ggtgctacgc ctgaataagt gataataagc ggatgaatgg     3060
cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc     3120
gcttatgtct attgctggtt tatcggtacc ccccaactga tcttcagcat cttttacttt     3180
caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag     3240
ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta     3300
tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaaa     3360
gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc     3420
tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca     3480
aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa     3540
aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct     3600
actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc     3660
ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga     3720
ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa     3780
cagccaagct tagcggccgc ttagaccttt cgctttttct tgggatcggc tctggagtcg     3840
ccaccaagct gagacaggtc gattcgggtc tcgtacaggc cagtgatgga ctggtgaatc     3900
agggtggcat cgagaacctc cttggtggat gtgtaccgct ttcggtcgat agtggtatcg     3960
aagtacttga aagctgcagg agcacccagg ttggtaagag taaacaggtg aatgatgttc     4020
tccgcctgtt ctcgaatggg tttgtcccga tgcttgttgt aggcagagag caccttgtcc     4080
aagttggcat cagccaggat gactcgcttc gaaaactcgg aaatctgctc gataatctcg     4140
tcgaggtaat gtttgtgctg ctcaacgaag agttgcttct gttcgttgtc ctcgggagaa     4200
cccttgagct tctcgtagtg agaagccaga tagagaaagt tgacgtactt cgaaggcaag     4260
gcaagctcgt ttcccttctg cagctcgcca gcggaggcga gcatacgctt tcgaccgttc     4320
tccagttcga acagagagta cttgggcagc ttgataatga ggtctttctt gacctccttg     4380
taacccttgg cttccaagaa gtcgatggga ttcttctcga agctcgatcg ctccatgatg     4440
gtaattccga gcagctcctt gacggacttg agctttttgg acttgccctt ctcgaccttc     4500
gcaacgacaa gcacggaata ggcgacggta ggagaatcga agccaccgta tttcttggga     4560
tcccagtctt tctttcgagc gatgagcttg tcggagtttc gcttgggcag aatcgactcc     4620
ttggagaatc cgccagtctg aacctcggtt ttcttgacga tgttgacctg aggcatcgac     4680
agaacctttc gcacggttgc aaagtctcga cccttgtccc acacgatctc tccagtttcg     4740
ccgttggtct cgataagtgg tctctttcga atctctccgt tggccaaggt gatctcggtc     4800
ttgaaaaagt tcatgatgtt ggagtaaaag aagtacttgg cagtagcctt gccaatctcc     4860
tgttcggact tggcaatcat ctttcgaacg tcgtagacct tgtaatcgcc gtaaacgaac     4920
tcgctttcga gcttggggta tttcttgatg agcgcagtgc caacgacggc gttgaggtaa     4980
gcatcgtggg catggtggta attgttgatc tctcgcacct tgtagaactg aaagtccttt     5040
cggaaatcgg agaccagttt ggacttgaga gtaatcacct tgacctctcg gatgagcttg     5100
tcgttctcgt cgtacttggt gttcatccga gaatcgagaa tctgtgcgac gtgctttgtg     5160
atctgtctgg tctcgacgag ttgacgcttg atgaagccag ccttgtcgag ctcggacaga     5220
ccgcctcgct cggccttggt aagattgtcg aactttcgct gggtaatgag cttggcgttg     5280
agcagctgtc gccagtagtt cttcatcttt ttgaccacct cttcgctggg aacgttgtcc     5340
gacttgcctc tgttcttgtc ggatcgtgta aggaccttgt tgtcgataga atcgtccttg     5400
agaaaggatt gagggacaat gtggtccaca tcgtagtcgc tgagacgatt gatgtccagt     5460
tcctgatcca cgtacatgtc tcgaccattc tgcagatagt agagatacag cttctcgttc     5520
tgcagttgag tgttctcgac gggatgctcc ttgagaatct gggatcccag ctccttgatg     5580
ccttcctcga ttcgcttcat ccgctctcgc gagtttttct gacccttttg agttgtctgg     5640
ttctctctgg ccatctcgat cacaatgttc tcgggcttgt gacgtcccat gaccttcacc     5700
agctcgtcga caaccttgac agtctggaga atgcctttct tgatggctgg cgaaccagcc     5760
aggttggcaa tatgttcgtg caagctgtcg ccctgaccgg acacttgtgc cttctggatg     5820
tcctccttga aggtaagaga atcgtcgtga atgagctgca tgaagtttcg gttggcaaag     5880
ccatcggact tgagaaagtc cagaatggtc tttccggact gcttgtctct gatgccgttg     5940
atgagctttc gcgaaagtct tccccagccg gtgtatctac gtcgcttgag ttgtttcatg     6000
accttgtcgt cgaacaggtg agcgtatgtc ttgagtcgtt cctcgatcat ctcccgatct     6060
tcgaacaggg taagagtgag cacgatgtcc tccagaatgt cctcgttttc ctcgttgtcg     6120
agaaaatcct tgtccttgat aatcttgagc agatcgtgat aggtgcccaa agaggcgttg     6180
aatcggtcct caactccgga aatctcgacg ctgtcgaaac actcgatttt cttgaagtag     6240
tcctccttga gctgcttaac agtgaccttt cggttggtct tgaacaggag atcgacaatg     6300
gctttcttct gttcgccaga caagaaggca ggctttcgca ttccctcggt aacgtacttg     6360
actttggtga gttcgttgta gactgtaaag tactcgtaga gcagcgaatg cttgggaaga     6420
accttctcgt tgggcagatt cttgtcgaag ttggtcattc gctcgatgaa ggactgtgca     6480
gaggcaccct tgtccacgac ttcctcgaag ttccagggag tgatggtttc ctcggacttt     6540
cgagtcatcc aagcaaatcg agagtttcct ctggcaagag gaccaacata gtaggggatt     6600
cgaaaggtaa gaatcttctc gatcttctct cggttgtcct tgagaaaggg gtagaagtct     6660
tcctgacgtc gaagaatggc gtgcagctca ccgaggtgga tctgatgagg aatgctgccg     6720
ttgtcgaagg ttcgttgctt ccgaagcaga tcctctcgat tgagcttgac aagcagttcc     6780
tcggttccgt ccatcttctc gagaattggc ttgatgaact tgtagaactc ttcctgagag     6840
gctccgccgt cgatgtatcc agcgtagccg ttcttcgact gatcgaaaaa gatctccttg     6900
tacttctcgg gcagttgctg tcggacaaga gccttgagca gtgtgagatc ctgatggtgc     6960
tcgtcgtatc gcttgatcat ggaggcagaa aggggagcct ttgtgatctc ggtgttgact     7020
cgcagaatgt cagacaagag aatagcatcc gaaaggttct tggcagcgag aaacaggtcg     7080
gcgtactgat cgccaatctg tgcaagcagg ttgtcgaggt catcgtcgta ggtgtccttg     7140
gacagctgga gcttggcgtc ctccgccaga tcgaagttgg acttgaagtt gggtgtgaga     7200
ccaagagaaa gggcaatgag gttgccaaac agtccgttct ttttctcgcc aggaagttgg     7260
gcaatgaggt tctccagtcg tctgctcttc gagagtcgag cagacaagat ggcctttgca     7320
tcgactccgg aggcattgat ggggttttcc tcgaacagct ggttgtaggt ctgaacgagc     7380
tgaatgaaca gcttgtccac atcgctgttg tcgggattga gatcgccctc gatgaggaaa     7440
tgacctcgaa acttgatcat gtgtgccaga gcgagatgga taagtctgag atccgccttg     7500
tcggtggaat cgacgagttt ctttcgcagg tggtagatgg taggatactt ctcgtggtaa     7560
gcaacctcgt ccacaatgtt gccaaagatg ggatgacgct cgtgtttctt gtcttcctcg     7620
acgaggaagg attcctccag tcgatgaaag aacgaatcgt ccaccttggc catctcgttg     7680
gaaaagatct cctgcaggta gcagattcgg ttcttccgtc gggtgtaacg tcgccgagca     7740
gttcgcttga gtctggtagc ttcggcagtc tcgccagaat cgaacaacag ggcaccaatg     7800
aggtttttct tgatggagtg tcgatcggtg tttccgagga ccttgaattt cttggagggc     7860
accttgtact cgtcggtgat gacagcccag ccgacagagt tggttccaat gtccaggccg     7920
atggagtatt tcttgtcgaa ttcccatatg gtaccagctg cagatctcga gctcggatcc     7980
ttatcgtcat cgtcgtacag atcccgaccc atttgctgtc caccagtcat gctagccata     8040
ccatgatgat gatgatgatg agaacccccc atggttaatt cctcctgtta gcccaaaaaa     8100
cgggtatgga gaaacagtag agagttgcga taaaaagcgt caggtaggat ccgctaatct     8160
tatggataaa aatgctatgg catagcaaag tgtgacgccg tgcaaataat caatgtggac     8220
ttttctgccg tgattataga cacttttgtt acgcgttttt gtcatggctt tggtcccgct     8280
ttgttacaga atgcttttaa taagcggggt taccggtttg gttagcgaga agagccagta     8340
aaagacgcag tgacggcaat gtctgatgca atatggacaa ttggtttctt ctctgaatgg     8400
cgggagtatg aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt     8460
taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc tcgatttttt     8520
tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc gcggtcaggg     8580
ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt tgctgttccc     8640
gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt atcaccagtg     8700
ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt caatatttgc     8760
caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg acctgtttgg     8820
gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg cgataaatct     8880
gcttgagcaa ttgttactgc ggcgcatgga agcgattaac gagtcgctcc atccaccgat     8940
ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag acagcaattt     9000
tgatatcgcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt     9060
ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc     9120
gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca atgttggttt     9180
tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca gcccgagcga     9240
gttccgtgcc ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt cataattggt     9300
aacgaatcag acaattgacg gc                                              9322

<210>  111
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  111
ggtttattga ctaccggaag c                                                 21

<210>  112
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  112
gccgtcaatt gtctgattcg                                                   20

<210>  113
<211>  10569
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF97-nacETsite1 plasmid

<400>  113
ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc       60
tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga      120
tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt      180
ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg      240
atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc      300
agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac      360
cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat      420
gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag      480
tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag      540
tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta      600
cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc      660
caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt      720
cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct      780
ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga      840
attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg      900
gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtttatc ggtacccccc aactgatctt cagcatcttt tactttcacc     1080
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg     1140
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag     1200
ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt     1260
ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc     1320
agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc     1380
cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg     1440
aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc     1500
tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca     1560
tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc     1620
ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc     1680
caagcttagc ggccgcttag acctttcgct ttttcttggg atcggctctg gagtcgccac     1740
caagctgaga caggtcgatt cgggtctcgt acaggccagt gatggactgg tgaatcaggg     1800
tggcatcgag aacctccttg gtggatgtgt accgctttcg gtcgatagtg gtatcgaagt     1860
acttgaaagc tgcaggagca cccaggttgg taagagtaaa caggtgaatg atgttctccg     1920
cctgttctcg aatgggtttg tcccgatgct tgttgtaggc agagagcacc ttgtccaagt     1980
tggcatcagc caggatgact cgcttcgaaa actcggaaat ctgctcgata atctcgtcga     2040
ggtaatgttt gtgctgctca acgaagagtt gcttctgttc gttgtcctcg ggagaaccct     2100
tgagcttctc gtagtgagaa gccagataga gaaagttgac gtacttcgaa ggcaaggcaa     2160
gctcgtttcc cttctgcagc tcgccagcgg aggcgagcat acgctttcga ccgttctcca     2220
gttcgaacag agagtacttg ggcagcttga taatgaggtc tttcttgacc tccttgtaac     2280
ccttggcttc caagaagtcg atgggattct tctcgaagct cgatcgctcc atgatggtaa     2340
ttccgagcag ctccttgacg gacttgagct ttttggactt gcccttctcg accttcgcaa     2400
cgacaagcac ggaataggcg acggtaggag aatcgaagcc accgtatttc ttgggatccc     2460
agtctttctt tcgagcgatg agcttgtcgg agtttcgctt gggcagaatc gactccttgg     2520
agaatccgcc agtctgaacc tcggttttct tgacgatgtt gacctgaggc atcgacagaa     2580
cctttcgcac ggttgcaaag tctcgaccct tgtcccacac gatctctcca gtttcgccgt     2640
tggtctcgat aagtggtctc tttcgaatct ctccgttggc caaggtgatc tcggtcttga     2700
aaaagttcat gatgttggag taaaagaagt acttggcagt agccttgcca atctcctgtt     2760
cggacttggc aatcatcttt cgaacgtcgt agaccttgta atcgccgtaa acgaactcgc     2820
tttcgagctt ggggtatttc ttgatgagcg cagtgccaac gacggcgttg aggtaagcat     2880
cgtgggcatg gtggtaattg ttgatctctc gcaccttgta gaactgaaag tcctttcgga     2940
aatcggagac cagtttggac ttgagagtaa tcaccttgac ctctcggatg agcttgtcgt     3000
tctcgtcgta cttggtgttc atccgagaat cgagaatctg tgcgacgtgc tttgtgatct     3060
gtctggtctc gacgagttga cgcttgatga agccagcctt gtcgagctcg gacagaccgc     3120
ctcgctcggc cttggtaaga ttgtcgaact ttcgctgggt aatgagcttg gcgttgagca     3180
gctgtcgcca gtagttcttc atctttttga ccacctcttc gctgggaacg ttgtccgact     3240
tgcctctgtt cttgtcggat cgtgtaagga ccttgttgtc gatagaatcg tccttgagaa     3300
aggattgagg gacaatgtgg tccacatcgt agtcgctgag acgattgatg tccagttcct     3360
gatccacgta catgtctcga ccattctgca gatagtagag atacagcttc tcgttctgca     3420
gttgagtgtt ctcgacggga tgctccttga gaatctggga tcccagctcc ttgatgcctt     3480
cctcgattcg cttcatccgc tctcgcgagt ttttctgacc cttttgagtt gtctggttct     3540
ctctggccat ctcgatcaca atgttctcgg gcttgtgacg tcccatgacc ttcaccagct     3600
cgtcgacaac cttgacagtc tggagaatgc ctttcttgat ggctggcgaa ccagccaggt     3660
tggcaatatg ttcgtgcaag ctgtcgccct gaccggacac ttgtgccttc tggatgtcct     3720
ccttgaaggt aagagaatcg tcgtgaatga gctgcatgaa gtttcggttg gcaaagccat     3780
cggacttgag aaagtccaga atggtctttc cggactgctt gtctctgatg ccgttgatga     3840
gctttcgcga aagtcttccc cagccggtgt atctacgtcg cttgagttgt ttcatgacct     3900
tgtcgtcgaa caggtgagcg tatgtcttga gtcgttcctc gatcatctcc cgatcttcga     3960
acagggtaag agtgagcacg atgtcctcca gaatgtcctc gttttcctcg ttgtcgagaa     4020
aatccttgtc cttgataatc ttgagcagat cgtgataggt gcccaaagag gcgttgaatc     4080
ggtcctcaac tccggaaatc tcgacgctgt cgaaacactc gattttcttg aagtagtcct     4140
ccttgagctg cttaacagtg acctttcggt tggtcttgaa caggagatcg acaatggctt     4200
tcttctgttc gccagacaag aaggcaggct ttcgcattcc ctcggtaacg tacttgactt     4260
tggtgagttc gttgtagact gtaaagtact cgtagagcag cgaatgcttg ggaagaacct     4320
tctcgttggg cagattcttg tcgaagttgg tcattcgctc gatgaaggac tgtgcagagg     4380
cacccttgtc cacgacttcc tcgaagttcc agggagtgat ggtttcctcg gactttcgag     4440
tcatccaagc aaatcgagag tttcctctgg caagaggacc aacatagtag gggattcgaa     4500
aggtaagaat cttctcgatc ttctctcggt tgtccttgag aaaggggtag aagtcttcct     4560
gacgtcgaag aatggcgtgc agctcaccga ggtggatctg atgaggaatg ctgccgttgt     4620
cgaaggttcg ttgcttccga agcagatcct ctcgattgag cttgacaagc agttcctcgg     4680
ttccgtccat cttctcgaga attggcttga tgaacttgta gaactcttcc tgagaggctc     4740
cgccgtcgat gtatccagcg tagccgttct tcgactgatc gaaaaagatc tccttgtact     4800
tctcgggcag ttgctgtcgg acaagagcct tgagcagtgt gagatcctga tggtgctcgt     4860
cgtatcgctt gatcatggag gcagaaaggg gagcctttgt gatctcggtg ttgactcgca     4920
gaatgtcaga caagagaata gcatccgaaa ggttcttggc agcgagaaac aggtcggcgt     4980
actgatcgcc aatctgtgca agcaggttgt cgaggtcatc gtcgtaggtg tccttggaca     5040
gctggagctt ggcgtcctcc gccagatcga agttggactt gaagttgggt gtgagaccaa     5100
gagaaagggc aatgaggttg ccaaacagtc cgttcttttt ctcgccagga agttgggcaa     5160
tgaggttctc cagtcgtctg ctcttcgaga gtcgagcaga caagatggcc tttgcatcga     5220
ctccggaggc attgatgggg ttttcctcga acagctggtt gtaggtctga acgagctgaa     5280
tgaacagctt gtccacatcg ctgttgtcgg gattgagatc gccctcgatg aggaaatgac     5340
ctcgaaactt gatcatgtgt gccagagcga ggtagataag tctgagatcc gccttgtcgg     5400
tggaatcgac gagtttcttt cgcaggtggt agatggtagg atacttctcg tggtaagcaa     5460
cctcgtccac aatgttgcca aagatgggat gacgctcgtg tttcttgtct tcctcgacga     5520
ggaaggattc ctccagtcga tgaaagaacg aatcgtccac cttggccatc tcgttggaaa     5580
agatctcctg caggtagcag attcggttct tccgtcgggt gtaacgtcgc cgagcagttc     5640
gcttgagtct ggtagcttcg gcagtctcgc cagaatcgaa caacagggca ccaatgaggt     5700
ttttcttgat ggagtgtcga tcggtgtttc cgaggacctt gaatttcttg gagggcacct     5760
tgtactcgtc ggtgatgaca gcccagccga cagagttggt tccaatgtcc aggccgatgg     5820
agtatttctt gtcgaattcc catatggtac cagctgcaga tctcgagctc ggatccttat     5880
cgtcatcgtc gtacagatcc cgacccattt gctgtccacc agtcatgcta gccataccat     5940
gatgatgatg atgatgagaa ccccccatgg ttaattcctc ctgttagccc aaaaaacggg     6000
tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg     6060
gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt     6120
ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt     6180
tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag     6240
acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg     6300
agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat     6360
gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc     6420
gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg     6480
gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca     6540
ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt     6600
tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat     6660
acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa     6720
atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt     6780
gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat     6840
aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat     6900
atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc     6960
cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag     7020
ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac     7080
gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc     7140
cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg     7200
aatcagacaa ttgacggccg tcccctggat tagctcgagc cgaacctccg ggaaaagttc     7260
gcgaaaagct ttaatgacct ctggcaagct ataacgtgcc tgagtatgcg tcgttgcaat     7320
agtgagaacg ccagacgtat cgttggtaaa caggtctgca agccgacgaa cattactggc     7380
ttcattcaga atacgttctg caatgaccag taatgctttg cccggttcag tcatgcccag     7440
cagtcgctta cctcgtcgaa caaatatttc gatgccaagt tcatcctcca gttcccgaat     7500
atgacggctg acgcctgact gtgaggtaaa aagcatattc gcaacctctg tcaggttgta     7560
atcctgacgt gcagcctcgc ggattatctt tagttgttgg aaattcacgg taaactccgg     7620
gcagttcaga tttcccgtta ttgttaaagt ctaatgcccg gcataacaaa taataaaaac     7680
ccgcatctta ttccatcccg atataacact tagctcacga agttcatgtt gcctccggtt     7740
tttaagaatc ggcccaagtg ccgccattac ttacaaccag attgcaagat gcttgccagt     7800
tttattttgg tgttgatgta caagctaacc aactgtcaaa taagagatta tgatagattc     7860
gtcatttgct cctttaatca gctgtcgcgt tcccctgccc tataaaagga gggtatgcac     7920
cacgatggtt cattacccaa taagattgaa agctcaccac tttgttgaaa ttgacagcaa     7980
acaaacaaaa aaatgcattt caccctttga catcaccatg cactgccatt aatatgcgcc     8040
ccgttcacac gattcctctg tagttcagtc ggtagaacgg cggactgtta atccgtatgt     8100
cactggttcg agtccagtca gaggagccaa attcaaaaaa gcctgctttc tagcaggctt     8160
tttgctttct aattaccaac gctcttaaaa catctgtctt gaaccagaac taatttgcaa     8220
gaatcataaa aaatttattt gctttcagga aaatttttct gtataataga ttcatatgca     8280
atacttcagc agccgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat     8340
caacttgaaa aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct     8400
cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg     8460
agaatggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctcaggct     8520
gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag     8580
ttgctttgtt ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac     8640
attgtcgatc tgttcatggt gaacagcttt aaatgcacca aaaactcgta aaagctctga     8700
tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat     8760
ctaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac     8820
aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc     8880
gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt tgcatgtcac     8940
tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg     9000
tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac     9060
cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt     9120
caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat     9180
tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa     9240
actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta     9300
tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca     9360
tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta     9420
actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga     9480
acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt     9540
ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc     9600
tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc     9660
ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt     9720
catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt     9780
cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta     9840
gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga     9900
cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt     9960
gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa    10020
agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt    10080
attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta    10140
aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg    10200
accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct    10260
ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa    10320
ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg    10380
ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc    10440
tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc    10500
agcggtatca tcaacaggct tacccgtctt actgtcggat cgacgctctc ccttatgcga    10560
ctcctgcat                                                            10569

<210>  114
<211>  10569
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF97-nacETsite2 plasmid

<400>  114
ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc       60
tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga      120
tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt      180
ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg      240
atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc      300
agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac      360
cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat      420
gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag      480
tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag      540
tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta      600
cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc      660
caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt      720
cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct      780
ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga      840
attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg      900
gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtttatc ggtacccccc aactgatctt cagcatcttt tactttcacc     1080
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg     1140
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag     1200
ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt     1260
ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc     1320
agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc     1380
cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg     1440
aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc     1500
tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca     1560
tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc     1620
ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc     1680
caagcttagc ggccgcttag acctttcgct ttttcttggg atcggctctg gagtcgccac     1740
caagctgaga caggtcgatt cgggtctcgt acaggccagt gatggactgg tgaatcaggg     1800
tggcatcgag aacctccttg gtggatgtgt accgctttcg gtcgatagtg gtatcgaagt     1860
acttgaaagc tgcaggagca cccaggttgg taagagtaaa caggtgaatg atgttctccg     1920
cctgttctcg aatgggtttg tcccgatgct tgttgtaggc agagagcacc ttgtccaagt     1980
tggcatcagc caggatgact cgcttcgaaa actcggaaat ctgctcgata atctcgtcga     2040
ggtaatgttt gtgctgctca acgaagagtt gcttctgttc gttgtcctcg ggagaaccct     2100
tgagcttctc gtagtgagaa gccagataga gaaagttgac gtacttcgaa ggcaaggcaa     2160
gctcgtttcc cttctgcagc tcgccagcgg aggcgagcat acgctttcga ccgttctcca     2220
gttcgaacag agagtacttg ggcagcttga taatgaggtc tttcttgacc tccttgtaac     2280
ccttggcttc caagaagtcg atgggattct tctcgaagct cgatcgctcc atgatggtaa     2340
ttccgagcag ctccttgacg gacttgagct ttttggactt gcccttctcg accttcgcaa     2400
cgacaagcac ggaataggcg acggtaggag aatcgaagcc accgtatttc ttgggatccc     2460
agtctttctt tcgagcgatg agcttgtcgg agtttcgctt gggcagaatc gactccttgg     2520
agaatccgcc agtctgaacc tcggttttct tgacgatgtt gacctgaggc atcgacagaa     2580
cctttcgcac ggttgcaaag tctcgaccct tgtcccacac gatctctcca gtttcgccgt     2640
tggtctcgat aagtggtctc tttcgaatct ctccgttggc caaggtgatc tcggtcttga     2700
aaaagttcat gatgttggag taaaagaagt acttggcagt agccttgcca atctcctgtt     2760
cggacttggc aatcatcttt cgaacgtcgt agaccttgta atcgccgtaa acgaactcgc     2820
tttcgagctt ggggtatttc ttgatgagcg cagtgccaac gacggcgttg aggtaagcat     2880
cgtgggcatg gtggtaattg ttgatctctc gcaccttgta gaactgaaag tcctttcgga     2940
aatcggagac cagtttggac ttgagagtaa tcaccttgac ctctcggatg agcttgtcgt     3000
tctcgtcgta cttggtgttc atccgagaat cgagaatctg tgcgacgtgc tttgtgatct     3060
gtctggtctc gacgagttga cgcttgatga agccagcctt gtcgagctcg gacagaccgc     3120
ctcgctcggc cttggtaaga ttgtcgaact ttcgctgggt aatgagcttg gcgttgagca     3180
gctgtcgcca gtagttcttc atctttttga ccacctcttc gctgggaacg ttgtccgact     3240
tgcctctgtt cttgtcggat cgtgtaagga ccttgttgtc gatagaatcg tccttgagaa     3300
aggattgagg gacaatgtgg tccacatcgt agtcgctgag acgattgatg tccagttcct     3360
gatccacgta catgtctcga ccattctgca gatagtagag atacagcttc tcgttctgca     3420
gttgagtgtt ctcgacggga tgctccttga gaatctggga tcccagctcc ttgatgcctt     3480
cctcgattcg cttcatccgc tctcgcgagt ttttctgacc cttttgagtt gtctggttct     3540
ctctggccat ctcgatcaca atgttctcgg gcttgtgacg tcccatgacc ttcaccagct     3600
cgtcgacaac cttgacagtc tggagaatgc ctttcttgat ggctggcgaa ccagccaggt     3660
tggcaatatg ttcgtgcaag ctgtcgccct gaccggacac ttgtgccttc tggatgtcct     3720
ccttgaaggt aagagaatcg tcgtgaatga gctgcatgaa gtttcggttg gcaaagccat     3780
cggacttgag aaagtccaga atggtctttc cggactgctt gtctctgatg ccgttgatga     3840
gctttcgcga aagtcttccc cagccggtgt atctacgtcg cttgagttgt ttcatgacct     3900
tgtcgtcgaa caggtgagcg tatgtcttga gtcgttcctc gatcatctcc cgatcttcga     3960
acagggtaag agtgagcacg atgtcctcca gaatgtcctc gttttcctcg ttgtcgagaa     4020
aatccttgtc cttgataatc ttgagcagat cgtgataggt gcccaaagag gcgttgaatc     4080
ggtcctcaac tccggaaatc tcgacgctgt cgaaacactc gattttcttg aagtagtcct     4140
ccttgagctg cttaacagtg acctttcggt tggtcttgaa caggagatcg acaatggctt     4200
tcttctgttc gccagacaag aaggcaggct ttcgcattcc ctcggtaacg tacttgactt     4260
tggtgagttc gttgtagact gtaaagtact cgtagagcag cgaatgcttg ggaagaacct     4320
tctcgttggg cagattcttg tcgaagttgg tcattcgctc gatgaaggac tgtgcagagg     4380
cacccttgtc cacgacttcc tcgaagttcc agggagtgat ggtttcctcg gactttcgag     4440
tcatccaagc aaatcgagag tttcctctgg caagaggacc aacatagtag gggattcgaa     4500
aggtaagaat cttctcgatc ttctctcggt tgtccttgag aaaggggtag aagtcttcct     4560
gacgtcgaag aatggcgtgc agctcaccga ggtggatctg atgaggaatg ctgccgttgt     4620
cgaaggttcg ttgcttccga agcagatcct ctcgattgag cttgacaagc agttcctcgg     4680
ttccgtccat cttctcgaga attggcttga tgaacttgta gaactcttcc tgagaggctc     4740
cgccgtcgat gtatccagcg tagccgttct tcgactgatc gaaaaagatc tccttgtact     4800
tctcgggcag ttgctgtcgg acaagagcct tgagcagtgt gagatcctga tggtgctcgt     4860
cgtatcgctt gatcatggag gcagaaaggg gagcctttgt gatctcggtg ttgactcgca     4920
gaatgtcaga caagagaata gcatccgaaa ggttcttggc agcgagaaac aggtcggcgt     4980
actgatcgcc aatctgtgca agcaggttgt cgaggtcatc gtcgtaggtg tccttggaca     5040
gctggagctt ggcgtcctcc gccagatcga agttggactt gaagttgggt gtgagaccaa     5100
gagaaagggc aatgaggttg ccaaacagtc cgttcttttt ctcgccagga agttgggcaa     5160
tgaggttctc cagtcgtctg ctcttcgaga gtcgagcaga caagatggcc tttgcatcga     5220
ctccggaggc attgatgggg ttttcctcga acagctggtt gtaggtctga acgagctgaa     5280
tgaacagctt gtccacatcg ctgttgtcgg gattgagatc gccctcgatg aggaaatgac     5340
ctcgaaactt gatcatgtgt gccagagcga ggtagataag tctgagatcc gccttgtcgg     5400
tggaatcgac gagtttcttt cgcaggtggt agatggtagg atacttctcg tggtaagcaa     5460
cctcgtccac aatgttgcca aagatgggat gacgctcgtg tttcttgtct tcctcgacga     5520
ggaaggattc ctccagtcga tgaaagaacg aatcgtccac cttggccatc tcgttggaaa     5580
agatctcctg caggtagcag attcggttct tccgtcgggt gtaacgtcgc cgagcagttc     5640
gcttgagtct ggtagcttcg gcagtctcgc cagaatcgaa caacagggca ccaatgaggt     5700
ttttcttgat ggagtgtcga tcggtgtttc cgaggacctt gaatttcttg gagggcacct     5760
tgtactcgtc ggtgatgaca gcccagccga cagagttggt tccaatgtcc aggccgatgg     5820
agtatttctt gtcgaattcc catatggtac cagctgcaga tctcgagctc ggatccttat     5880
cgtcatcgtc gtacagatcc cgacccattt gctgtccacc agtcatgcta gccataccat     5940
gatgatgatg atgatgagaa ccccccatgg ttaattcctc ctgttagccc aaaaaacggg     6000
tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg     6060
gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt     6120
ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt     6180
tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag     6240
acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg     6300
agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat     6360
gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc     6420
gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg     6480
gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca     6540
ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt     6600
tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat     6660
acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa     6720
atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt     6780
gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat     6840
aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat     6900
atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc     6960
cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag     7020
ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac     7080
gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc     7140
cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg     7200
aatcagacaa ttgacggccg tcccctggat tagctcgagc cgaacctccg ggaaaagttc     7260
gcgaaaagct ttaatgacct ctggcaagct ataacgtgcc tgagtatgcg tcgttgcaat     7320
agtgagaacg ccagacgtat cgttggtaaa caggtctgca agccgacgaa cattactggc     7380
ttcattcaga atacgttctg caatgaccag taatgctttg cccggttcag tcatgcccag     7440
cagtcgctta cctcgtcgaa caaatatttc gatgccaagt tcatcctcca gttcccgaat     7500
atgacggctg acgcctgact gtgaggtaaa aagcatattc gcaacctctg tcaggttgta     7560
atcctgacgt gcagcctcgc ggattatctt tagttgttgg aaattcacgg taaactccgg     7620
gcagttcaga tttcccgtta ttgttaaagt ctaatgcccg gcataacaaa taataaaaac     7680
ccgcatctta ttccatcccg atataacact tagctcacga agttcatgtt gcctccggtt     7740
tttaagaatc ggcccaagtg ccgccattac ttacaaccag attgcaagat gcttgccagt     7800
tttattttgg tgttgatgta caagctaacc aactgtcaaa taagagatta tgatagattc     7860
gtcatttgct cctttaatca gctgtcgcgt tcccctgccc tataaaagga gggtatgcac     7920
cacgatggtt cattacccaa taagattgaa agctcaccac tttgttgaaa ttgacagcaa     7980
acaaacaaaa aaatgcattt caccctttga catcaccatg cactgccatt aatatgcgcc     8040
ccgttcacac gattcctctg tagttcagtc ggtagaacgg cggactgtta atccgtatgt     8100
cactggttcg agtccagtca gaggagccaa attcaaaaaa gcctgctttc tagcaggctt     8160
tttgctttct aattaccaac gctcttaaaa catctgtctt gaaccagaac taatttgcaa     8220
gaatcataaa aaatttattt gctttcagga aaatttttct gtataataga ttcaacaacc     8280
agcgctcagc cagcgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat     8340
caacttgaaa aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct     8400
cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg     8460
agaatggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctcaggct     8520
gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag     8580
ttgctttgtt ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac     8640
attgtcgatc tgttcatggt gaacagcttt aaatgcacca aaaactcgta aaagctctga     8700
tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat     8760
ctaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac     8820
aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc     8880
gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt tgcatgtcac     8940
tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg     9000
tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac     9060
cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt     9120
caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat     9180
tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa     9240
actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta     9300
tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca     9360
tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta     9420
actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga     9480
acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt     9540
ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc     9600
tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc     9660
ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt     9720
catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt     9780
cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta     9840
gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga     9900
cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt     9960
gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa    10020
agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt    10080
attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta    10140
aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg    10200
accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct    10260
ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa    10320
ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg    10380
ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc    10440
tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc    10500
agcggtatca tcaacaggct tacccgtctt actgtcggat cgacgctctc ccttatgcga    10560
ctcctgcat                                                            10569

<210>  115
<211>  10569
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pRF861-nacETsite1 plasmid

<400>  115
ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc       60
tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga      120
tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt      180
ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg      240
atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc      300
agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac      360
cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat      420
gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag      480
tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag      540
tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta      600
cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc      660
caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt      720
cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct      780
ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga      840
attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg      900
gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtttatc ggtacccccc aactgatctt cagcatcttt tactttcacc     1080
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg     1140
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag     1200
ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt     1260
ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc     1320
agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc     1380
cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg     1440
aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc     1500
tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca     1560
tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc     1620
ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc     1680
caagcttagc ggccgcttag acctttcgct ttttcttggg atcggctctg gagtcgccac     1740
caagctgaga caggtcgatt cgggtctcgt acaggccagt gatggactgg tgaatcaggg     1800
tggcatcgag aacctccttg gtggatgtgt accgctttcg gtcgatagtg gtatcgaagt     1860
acttgaaagc tgcaggagca cccaggttgg taagagtaaa caggtgaatg atgttctccg     1920
cctgttctcg aatgggtttg tcccgatgct tgttgtaggc agagagcacc ttgtccaagt     1980
tggcatcagc caggatgact cgcttcgaaa actcggaaat ctgctcgata atctcgtcga     2040
ggtaatgttt gtgctgctca acgaagagtt gcttctgttc gttgtcctcg ggagaaccct     2100
tgagcttctc gtagtgagaa gccagataga gaaagttgac gtacttcgaa ggcaaggcaa     2160
gctcgtttcc cttctgcagc tcgccagcgg aggcgagcat acgctttcga ccgttctcca     2220
gttcgaacag agagtacttg ggcagcttga taatgaggtc tttcttgacc tccttgtaac     2280
ccttggcttc caagaagtcg atgggattct tctcgaagct cgatcgctcc atgatggtaa     2340
ttccgagcag ctccttgacg gacttgagct ttttggactt gcccttctcg accttcgcaa     2400
cgacaagcac ggaataggcg acggtaggag aatcgaagcc accgtatttc ttgggatccc     2460
agtctttctt tcgagcgatg agcttgtcgg agtttcgctt gggcagaatc gactccttgg     2520
agaatccgcc agtctgaacc tcggttttct tgacgatgtt gacctgaggc atcgacagaa     2580
cctttcgcac ggttgcaaag tctcgaccct tgtcccacac gatctctcca gtttcgccgt     2640
tggtctcgat aagtggtctc tttcgaatct ctccgttggc caaggtgatc tcggtcttga     2700
aaaagttcat gatgttggag taaaagaagt acttggcagt agccttgcca atctcctgtt     2760
cggacttggc aatcatcttt cgaacgtcgt agaccttgta atcgccgtaa acgaactcgc     2820
tttcgagctt ggggtatttc ttgatgagcg cagtgccaac gacggcgttg aggtaagcat     2880
cgtgggcatg gtggtaattg ttgatctctc gcaccttgta gaactgaaag tcctttcgga     2940
aatcggagac cagtttggac ttgagagtaa tcaccttgac ctctcggatg agcttgtcgt     3000
tctcgtcgta cttggtgttc atccgagaat cgagaatctg tgcgacgtgc tttgtgatct     3060
gtctggtctc gacgagttga cgcttgatga agccagcctt gtcgagctcg gacagaccgc     3120
ctcgctcggc cttggtaaga ttgtcgaact ttcgctgggt aatgagcttg gcgttgagca     3180
gctgtcgcca gtagttcttc atctttttga ccacctcttc gctgggaacg ttgtccgact     3240
tgcctctgtt cttgtcggat cgtgtaagga ccttgttgtc gatagaatcg tccttgagaa     3300
aggattgagg gacaatgtgg tccacatcgt agtcgctgag acgattgatg tccagttcct     3360
gatccacgta catgtctcga ccattctgca gatagtagag atacagcttc tcgttctgca     3420
gttgagtgtt ctcgacggga tgctccttga gaatctggga tcccagctcc ttgatgcctt     3480
cctcgattcg cttcatccgc tctcgcgagt ttttctgacc cttttgagtt gtctggttct     3540
ctctggccat ctcgatcaca atgttctcgg gcttgtgacg tcccatgacc ttcaccagct     3600
cgtcgacaac cttgacagtc tggagaatgc ctttcttgat ggctggcgaa ccagccaggt     3660
tggcaatatg ttcgtgcaag ctgtcgccct gaccggacac ttgtgccttc tggatgtcct     3720
ccttgaaggt aagagaatcg tcgtgaatga gctgcatgaa gtttcggttg gcaaagccat     3780
cggacttgag aaagtccaga atggtctttc cggactgctt gtctctgatg ccgttgatga     3840
gctttcgcga aagtcttccc cagccggtgt atctacgtcg cttgagttgt ttcatgacct     3900
tgtcgtcgaa caggtgagcg tatgtcttga gtcgttcctc gatcatctcc cgatcttcga     3960
acagggtaag agtgagcacg atgtcctcca gaatgtcctc gttttcctcg ttgtcgagaa     4020
aatccttgtc cttgataatc ttgagcagat cgtgataggt gcccaaagag gcgttgaatc     4080
ggtcctcaac tccggaaatc tcgacgctgt cgaaacactc gattttcttg aagtagtcct     4140
ccttgagctg cttaacagtg acctttcggt tggtcttgaa caggagatcg acaatggctt     4200
tcttctgttc gccagacaag aaggcaggct ttcgcattcc ctcggtaacg tacttgactt     4260
tggtgagttc gttgtagact gtaaagtact cgtagagcag cgaatgcttg ggaagaacct     4320
tctcgttggg cagattcttg tcgaagttgg tcattcgctc gatgaaggac tgtgcagagg     4380
cacccttgtc cacgacttcc tcgaagttcc agggagtgat ggtttcctcg gactttcgag     4440
tcatccaagc aaatcgagag tttcctctgg caagaggacc aacatagtag gggattcgaa     4500
aggtaagaat cttctcgatc ttctctcggt tgtccttgag aaaggggtag aagtcttcct     4560
gacgtcgaag aatggcgtgc agctcaccga ggtggatctg atgaggaatg ctgccgttgt     4620
cgaaggttcg ttgcttccga agcagatcct ctcgattgag cttgacaagc agttcctcgg     4680
ttccgtccat cttctcgaga attggcttga tgaacttgta gaactcttcc tgagaggctc     4740
cgccgtcgat gtatccagcg tagccgttct tcgactgatc gaaaaagatc tccttgtact     4800
tctcgggcag ttgctgtcgg acaagagcct tgagcagtgt gagatcctga tggtgctcgt     4860
cgtatcgctt gatcatggag gcagaaaggg gagcctttgt gatctcggtg ttgactcgca     4920
gaatgtcaga caagagaata gcatccgaaa ggttcttggc agcgagaaac aggtcggcgt     4980
actgatcgcc aatctgtgca agcaggttgt cgaggtcatc gtcgtaggtg tccttggaca     5040
gctggagctt ggcgtcctcc gccagatcga agttggactt gaagttgggt gtgagaccaa     5100
gagaaagggc aatgaggttg ccaaacagtc cgttcttttt ctcgccagga agttgggcaa     5160
tgaggttctc cagtcgtctg ctcttcgaga gtcgagcaga caagatggcc tttgcatcga     5220
ctccggaggc attgatgggg ttttcctcga acagctggtt gtaggtctga acgagctgaa     5280
tgaacagctt gtccacatcg ctgttgtcgg gattgagatc gccctcgatg aggaaatgac     5340
ctcgaaactt gatcatgtgt gccagagcga gatggataag tctgagatcc gccttgtcgg     5400
tggaatcgac gagtttcttt cgcaggtggt agatggtagg atacttctcg tggtaagcaa     5460
cctcgtccac aatgttgcca aagatgggat gacgctcgtg tttcttgtct tcctcgacga     5520
ggaaggattc ctccagtcga tgaaagaacg aatcgtccac cttggccatc tcgttggaaa     5580
agatctcctg caggtagcag attcggttct tccgtcgggt gtaacgtcgc cgagcagttc     5640
gcttgagtct ggtagcttcg gcagtctcgc cagaatcgaa caacagggca ccaatgaggt     5700
ttttcttgat ggagtgtcga tcggtgtttc cgaggacctt gaatttcttg gagggcacct     5760
tgtactcgtc ggtgatgaca gcccagccga cagagttggt tccaatgtcc aggccgatgg     5820
agtatttctt gtcgaattcc catatggtac cagctgcaga tctcgagctc ggatccttat     5880
cgtcatcgtc gtacagatcc cgacccattt gctgtccacc agtcatgcta gccataccat     5940
gatgatgatg atgatgagaa ccccccatgg ttaattcctc ctgttagccc aaaaaacggg     6000
tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg     6060
gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt     6120
ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt     6180
tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag     6240
acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg     6300
agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat     6360
gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc     6420
gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg     6480
gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca     6540
ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt     6600
tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat     6660
acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa     6720
atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt     6780
gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat     6840
aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat     6900
atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc     6960
cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag     7020
ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac     7080
gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc     7140
cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg     7200
aatcagacaa ttgacggccg tcccctggat tagctcgagc cgaacctccg ggaaaagttc     7260
gcgaaaagct ttaatgacct ctggcaagct ataacgtgcc tgagtatgcg tcgttgcaat     7320
agtgagaacg ccagacgtat cgttggtaaa caggtctgca agccgacgaa cattactggc     7380
ttcattcaga atacgttctg caatgaccag taatgctttg cccggttcag tcatgcccag     7440
cagtcgctta cctcgtcgaa caaatatttc gatgccaagt tcatcctcca gttcccgaat     7500
atgacggctg acgcctgact gtgaggtaaa aagcatattc gcaacctctg tcaggttgta     7560
atcctgacgt gcagcctcgc ggattatctt tagttgttgg aaattcacgg taaactccgg     7620
gcagttcaga tttcccgtta ttgttaaagt ctaatgcccg gcataacaaa taataaaaac     7680
ccgcatctta ttccatcccg atataacact tagctcacga agttcatgtt gcctccggtt     7740
tttaagaatc ggcccaagtg ccgccattac ttacaaccag attgcaagat gcttgccagt     7800
tttattttgg tgttgatgta caagctaacc aactgtcaaa taagagatta tgatagattc     7860
gtcatttgct cctttaatca gctgtcgcgt tcccctgccc tataaaagga gggtatgcac     7920
cacgatggtt cattacccaa taagattgaa agctcaccac tttgttgaaa ttgacagcaa     7980
acaaacaaaa aaatgcattt caccctttga catcaccatg cactgccatt aatatgcgcc     8040
ccgttcacac gattcctctg tagttcagtc ggtagaacgg cggactgtta atccgtatgt     8100
cactggttcg agtccagtca gaggagccaa attcaaaaaa gcctgctttc tagcaggctt     8160
tttgctttct aattaccaac gctcttaaaa catctgtctt gaaccagaac taatttgcaa     8220
gaatcataaa aaatttattt gctttcagga aaatttttct gtataataga ttcatatgca     8280
atacttcagc agccgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat     8340
caacttgaaa aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct     8400
cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg     8460
agaatggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctcaggct     8520
gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag     8580
ttgctttgtt ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac     8640
attgtcgatc tgttcatggt gaacagcttt aaatgcacca aaaactcgta aaagctctga     8700
tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat     8760
ctaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac     8820
aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc     8880
gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt tgcatgtcac     8940
tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg     9000
tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac     9060
cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt     9120
caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat     9180
tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa     9240
actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta     9300
tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca     9360
tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta     9420
actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga     9480
acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt     9540
ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc     9600
tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc     9660
ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt     9720
catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt     9780
cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta     9840
gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga     9900
cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt     9960
gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa    10020
agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt    10080
attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta    10140
aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg    10200
accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct    10260
ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa    10320
ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg    10380
ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc    10440
tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc    10500
agcggtatca tcaacaggct tacccgtctt actgtcggat cgacgctctc ccttatgcga    10560
ctcctgcat                                                            10569

<210>  116
<211>  10569
<212>  DNA
<213>  Artificial sequence

<220>
<223>   pRF861-nacETsite2 plasmid

<400>  116
ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc       60
tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga      120
tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt      180
ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg      240
atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc      300
agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac      360
cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat      420
gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag      480
tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag      540
tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta      600
cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc      660
caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt      720
cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct      780
ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga      840
attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg      900
gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga      960
aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020
atgtctattg ctggtttatc ggtacccccc aactgatctt cagcatcttt tactttcacc     1080
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg     1140
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag     1200
ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt     1260
ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc     1320
agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc     1380
cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg     1440
aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc     1500
tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca     1560
tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc     1620
ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc     1680
caagcttagc ggccgcttag acctttcgct ttttcttggg atcggctctg gagtcgccac     1740
caagctgaga caggtcgatt cgggtctcgt acaggccagt gatggactgg tgaatcaggg     1800
tggcatcgag aacctccttg gtggatgtgt accgctttcg gtcgatagtg gtatcgaagt     1860
acttgaaagc tgcaggagca cccaggttgg taagagtaaa caggtgaatg atgttctccg     1920
cctgttctcg aatgggtttg tcccgatgct tgttgtaggc agagagcacc ttgtccaagt     1980
tggcatcagc caggatgact cgcttcgaaa actcggaaat ctgctcgata atctcgtcga     2040
ggtaatgttt gtgctgctca acgaagagtt gcttctgttc gttgtcctcg ggagaaccct     2100
tgagcttctc gtagtgagaa gccagataga gaaagttgac gtacttcgaa ggcaaggcaa     2160
gctcgtttcc cttctgcagc tcgccagcgg aggcgagcat acgctttcga ccgttctcca     2220
gttcgaacag agagtacttg ggcagcttga taatgaggtc tttcttgacc tccttgtaac     2280
ccttggcttc caagaagtcg atgggattct tctcgaagct cgatcgctcc atgatggtaa     2340
ttccgagcag ctccttgacg gacttgagct ttttggactt gcccttctcg accttcgcaa     2400
cgacaagcac ggaataggcg acggtaggag aatcgaagcc accgtatttc ttgggatccc     2460
agtctttctt tcgagcgatg agcttgtcgg agtttcgctt gggcagaatc gactccttgg     2520
agaatccgcc agtctgaacc tcggttttct tgacgatgtt gacctgaggc atcgacagaa     2580
cctttcgcac ggttgcaaag tctcgaccct tgtcccacac gatctctcca gtttcgccgt     2640
tggtctcgat aagtggtctc tttcgaatct ctccgttggc caaggtgatc tcggtcttga     2700
aaaagttcat gatgttggag taaaagaagt acttggcagt agccttgcca atctcctgtt     2760
cggacttggc aatcatcttt cgaacgtcgt agaccttgta atcgccgtaa acgaactcgc     2820
tttcgagctt ggggtatttc ttgatgagcg cagtgccaac gacggcgttg aggtaagcat     2880
cgtgggcatg gtggtaattg ttgatctctc gcaccttgta gaactgaaag tcctttcgga     2940
aatcggagac cagtttggac ttgagagtaa tcaccttgac ctctcggatg agcttgtcgt     3000
tctcgtcgta cttggtgttc atccgagaat cgagaatctg tgcgacgtgc tttgtgatct     3060
gtctggtctc gacgagttga cgcttgatga agccagcctt gtcgagctcg gacagaccgc     3120
ctcgctcggc cttggtaaga ttgtcgaact ttcgctgggt aatgagcttg gcgttgagca     3180
gctgtcgcca gtagttcttc atctttttga ccacctcttc gctgggaacg ttgtccgact     3240
tgcctctgtt cttgtcggat cgtgtaagga ccttgttgtc gatagaatcg tccttgagaa     3300
aggattgagg gacaatgtgg tccacatcgt agtcgctgag acgattgatg tccagttcct     3360
gatccacgta catgtctcga ccattctgca gatagtagag atacagcttc tcgttctgca     3420
gttgagtgtt ctcgacggga tgctccttga gaatctggga tcccagctcc ttgatgcctt     3480
cctcgattcg cttcatccgc tctcgcgagt ttttctgacc cttttgagtt gtctggttct     3540
ctctggccat ctcgatcaca atgttctcgg gcttgtgacg tcccatgacc ttcaccagct     3600
cgtcgacaac cttgacagtc tggagaatgc ctttcttgat ggctggcgaa ccagccaggt     3660
tggcaatatg ttcgtgcaag ctgtcgccct gaccggacac ttgtgccttc tggatgtcct     3720
ccttgaaggt aagagaatcg tcgtgaatga gctgcatgaa gtttcggttg gcaaagccat     3780
cggacttgag aaagtccaga atggtctttc cggactgctt gtctctgatg ccgttgatga     3840
gctttcgcga aagtcttccc cagccggtgt atctacgtcg cttgagttgt ttcatgacct     3900
tgtcgtcgaa caggtgagcg tatgtcttga gtcgttcctc gatcatctcc cgatcttcga     3960
acagggtaag agtgagcacg atgtcctcca gaatgtcctc gttttcctcg ttgtcgagaa     4020
aatccttgtc cttgataatc ttgagcagat cgtgataggt gcccaaagag gcgttgaatc     4080
ggtcctcaac tccggaaatc tcgacgctgt cgaaacactc gattttcttg aagtagtcct     4140
ccttgagctg cttaacagtg acctttcggt tggtcttgaa caggagatcg acaatggctt     4200
tcttctgttc gccagacaag aaggcaggct ttcgcattcc ctcggtaacg tacttgactt     4260
tggtgagttc gttgtagact gtaaagtact cgtagagcag cgaatgcttg ggaagaacct     4320
tctcgttggg cagattcttg tcgaagttgg tcattcgctc gatgaaggac tgtgcagagg     4380
cacccttgtc cacgacttcc tcgaagttcc agggagtgat ggtttcctcg gactttcgag     4440
tcatccaagc aaatcgagag tttcctctgg caagaggacc aacatagtag gggattcgaa     4500
aggtaagaat cttctcgatc ttctctcggt tgtccttgag aaaggggtag aagtcttcct     4560
gacgtcgaag aatggcgtgc agctcaccga ggtggatctg atgaggaatg ctgccgttgt     4620
cgaaggttcg ttgcttccga agcagatcct ctcgattgag cttgacaagc agttcctcgg     4680
ttccgtccat cttctcgaga attggcttga tgaacttgta gaactcttcc tgagaggctc     4740
cgccgtcgat gtatccagcg tagccgttct tcgactgatc gaaaaagatc tccttgtact     4800
tctcgggcag ttgctgtcgg acaagagcct tgagcagtgt gagatcctga tggtgctcgt     4860
cgtatcgctt gatcatggag gcagaaaggg gagcctttgt gatctcggtg ttgactcgca     4920
gaatgtcaga caagagaata gcatccgaaa ggttcttggc agcgagaaac aggtcggcgt     4980
actgatcgcc aatctgtgca agcaggttgt cgaggtcatc gtcgtaggtg tccttggaca     5040
gctggagctt ggcgtcctcc gccagatcga agttggactt gaagttgggt gtgagaccaa     5100
gagaaagggc aatgaggttg ccaaacagtc cgttcttttt ctcgccagga agttgggcaa     5160
tgaggttctc cagtcgtctg ctcttcgaga gtcgagcaga caagatggcc tttgcatcga     5220
ctccggaggc attgatgggg ttttcctcga acagctggtt gtaggtctga acgagctgaa     5280
tgaacagctt gtccacatcg ctgttgtcgg gattgagatc gccctcgatg aggaaatgac     5340
ctcgaaactt gatcatgtgt gccagagcga gatggataag tctgagatcc gccttgtcgg     5400
tggaatcgac gagtttcttt cgcaggtggt agatggtagg atacttctcg tggtaagcaa     5460
cctcgtccac aatgttgcca aagatgggat gacgctcgtg tttcttgtct tcctcgacga     5520
ggaaggattc ctccagtcga tgaaagaacg aatcgtccac cttggccatc tcgttggaaa     5580
agatctcctg caggtagcag attcggttct tccgtcgggt gtaacgtcgc cgagcagttc     5640
gcttgagtct ggtagcttcg gcagtctcgc cagaatcgaa caacagggca ccaatgaggt     5700
ttttcttgat ggagtgtcga tcggtgtttc cgaggacctt gaatttcttg gagggcacct     5760
tgtactcgtc ggtgatgaca gcccagccga cagagttggt tccaatgtcc aggccgatgg     5820
agtatttctt gtcgaattcc catatggtac cagctgcaga tctcgagctc ggatccttat     5880
cgtcatcgtc gtacagatcc cgacccattt gctgtccacc agtcatgcta gccataccat     5940
gatgatgatg atgatgagaa ccccccatgg ttaattcctc ctgttagccc aaaaaacggg     6000
tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg     6060
gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt     6120
ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt     6180
tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag     6240
acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg     6300
agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat     6360
gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc     6420
gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg     6480
gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca     6540
ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt     6600
tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat     6660
acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa     6720
atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt     6780
gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat     6840
aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat     6900
atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc     6960
cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag     7020
ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac     7080
gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc     7140
cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg     7200
aatcagacaa ttgacggccg tcccctggat tagctcgagc cgaacctccg ggaaaagttc     7260
gcgaaaagct ttaatgacct ctggcaagct ataacgtgcc tgagtatgcg tcgttgcaat     7320
agtgagaacg ccagacgtat cgttggtaaa caggtctgca agccgacgaa cattactggc     7380
ttcattcaga atacgttctg caatgaccag taatgctttg cccggttcag tcatgcccag     7440
cagtcgctta cctcgtcgaa caaatatttc gatgccaagt tcatcctcca gttcccgaat     7500
atgacggctg acgcctgact gtgaggtaaa aagcatattc gcaacctctg tcaggttgta     7560
atcctgacgt gcagcctcgc ggattatctt tagttgttgg aaattcacgg taaactccgg     7620
gcagttcaga tttcccgtta ttgttaaagt ctaatgcccg gcataacaaa taataaaaac     7680
ccgcatctta ttccatcccg atataacact tagctcacga agttcatgtt gcctccggtt     7740
tttaagaatc ggcccaagtg ccgccattac ttacaaccag attgcaagat gcttgccagt     7800
tttattttgg tgttgatgta caagctaacc aactgtcaaa taagagatta tgatagattc     7860
gtcatttgct cctttaatca gctgtcgcgt tcccctgccc tataaaagga gggtatgcac     7920
cacgatggtt cattacccaa taagattgaa agctcaccac tttgttgaaa ttgacagcaa     7980
acaaacaaaa aaatgcattt caccctttga catcaccatg cactgccatt aatatgcgcc     8040
ccgttcacac gattcctctg tagttcagtc ggtagaacgg cggactgtta atccgtatgt     8100
cactggttcg agtccagtca gaggagccaa attcaaaaaa gcctgctttc tagcaggctt     8160
tttgctttct aattaccaac gctcttaaaa catctgtctt gaaccagaac taatttgcaa     8220
gaatcataaa aaatttattt gctttcagga aaatttttct gtataataga ttcaacaacc     8280
agcgctcagc cagcgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat     8340
caacttgaaa aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct     8400
cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg     8460
agaatggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctcaggct     8520
gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag     8580
ttgctttgtt ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac     8640
attgtcgatc tgttcatggt gaacagcttt aaatgcacca aaaactcgta aaagctctga     8700
tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat     8760
ctaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac     8820
aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc     8880
gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt tgcatgtcac     8940
tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg     9000
tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac     9060
cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt     9120
caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat     9180
tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa     9240
actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta     9300
tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca     9360
tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta     9420
actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga     9480
acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt     9540
ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc     9600
tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc     9660
ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt     9720
catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt     9780
cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta     9840
gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga     9900
cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt     9960
gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa    10020
agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt    10080
attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta    10140
aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg    10200
accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct    10260
ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa    10320
ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg    10380
ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc    10440
tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc    10500
agcggtatca tcaacaggct tacccgtctt actgtcggat cgacgctctc ccttatgcga    10560
ctcctgcat                                                            10569

<210>  117
<211>  1958
<212>  DNA
<213>  E. coli

<400>  117
caatgtcgca atttcctgtg gcgtcccctg gattagctcg agccgaacct ccgggaaaag       60
ttcgcgaaaa gctttaatga cctctggcaa gctataacgt gcctgagtat gcgtcgttgc      120
aatagtgaga acgccagacg tatcgttggt aaacaggtct gcaagccgac gaacattact      180
ggcttcattc agaatacgtt ctgcaatgac cagtaatgct ttgcccggtt cagtcatgcc      240
cagcagtcgc ttacctcgtc gaacaaatat ttcgatgcca agttcatcct ccagttcccg      300
aatatgacgg ctgacgcctg actgtgaggt aaaaagcata ttcgcaacct ctgtcaggtt      360
gtaatcctga cgtgcagcct cgcggattat ctttagttgt tggaaattca cggtaaactc      420
cgggcagttc agatttcccg ttattgttaa agtctaatgc ccggcataac aaataataaa      480
aacccgcatc ttattccatc ccgatataac acttagctca ccaattgcca ctgccttttt      540
tccatcactg gagaactaat cactgacatt aacaactctt tcactgcctg tgcctgtggc      600
gataagttcg ctctggcggg taaatttaat gacaaagaga gactcatgga aggagtggta      660
atgcgtgaca tccacccatt tactgcgcca cataacgaac gcgcggccga ttcgggtaat      720
actgcaacgc ccatgccgct ggcaatcgct gcggtaagcg tggcaataga ctcaatttca      780
ccaataactt ttgccgtgag tcgccgtagg gaaaaagcct catcaacacg aagtctaata      840
gcactgtaat cactggggag aaagaggttc atttgcgcaa tagcattcac atcaacgctt      900
tgccccgggc aatcttgagt tcctaccaga aaaagatctt ctttcagcaa agcctgactg      960
gatacaccag ccacagggga atgctcataa atcaccgcca tatcgagttg gtgatttatc     1020
aatttttcgt taagcactgc accactattt tcatgaagat agataacgat ctccggaaat     1080
tcagcgcgaa ccgcctgtaa taagggcatg gtgatggatg acgcagcggt tcctggtgca     1140
aagccaatcg agacttgccc cgataatgcc tgaccaacgt tatgcaccgc cagttgggcc     1200
tgttcacact gacgtaaaat ggcccgcgca tgggtataga gaatttttcc ggcgtctgtt     1260
ggtgtaacgc cccgctttgt acggatcaaa agttgttgat ttaactcacc ttccagtgtg     1320
gcaacctgct ggctgagcgc tggttgtgcg atatgcaata cttcagcagc ctgggtcagg     1380
ctaccaatat ctacaatttt tacgaagtat ttcaggcgtc tgaagttcat gttgcctccg     1440
gtttttaaga atcggcccaa gtgccgccat tacttacaac cagattgcaa gatgcttgcc     1500
agttttattt tggtgttgat gtacaagcta accaactgtc aaataagaga ttatgataga     1560
ttcgtcattt gctcctttaa tcagctgtcg cgttcccctg ccctataaaa ggagggtatg     1620
caccacgatg gttcattacc caataagatt gaaagctcac cactttgttg aaattgacag     1680
caaacaaaca aaaaaatgca tttcaccctt tgacatcacc atgcactgcc attaatatgc     1740
gccccgttca cacgattcct ctgtagttca gtcggtagaa cggcggactg ttaatccgta     1800
tgtcactggt tcgagtccag tcagaggagc caaattcaaa aaagcctgct ttctagcagg     1860
ctttttgctt tctaattacc aacgctctta aaacatctgt cttgaaccag aactaatttg     1920
cacaggcatt cccgatcgac gttgcaacgc agcatttg                             1958

<210>  118
<211>  1058
<212>  DNA
<213>  artificial sequence

<220>
<223>  edited nac locus

<400>  118
caatgtcgca atttcctgtg gcgtcccctg gattagctcg agccgaacct ccgggaaaag       60
ttcgcgaaaa gctttaatga cctctggcaa gctataacgt gcctgagtat gcgtcgttgc      120
aatagtgaga acgccagacg tatcgttggt aaacaggtct gcaagccgac gaacattact      180
ggcttcattc agaatacgtt ctgcaatgac cagtaatgct ttgcccggtt cagtcatgcc      240
cagcagtcgc ttacctcgtc gaacaaatat ttcgatgcca agttcatcct ccagttcccg      300
aatatgacgg ctgacgcctg actgtgaggt aaaaagcata ttcgcaacct ctgtcaggtt      360
gtaatcctga cgtgcagcct cgcggattat ctttagttgt tggaaattca cggtaaactc      420
cgggcagttc agatttcccg ttattgttaa agtctaatgc ccggcataac aaataataaa      480
aacccgcatc ttattccatc ccgatataac acttagctca cgaagttcat gttgcctccg      540
gtttttaaga atcggcccaa gtgccgccat tacttacaac cagattgcaa gatgcttgcc      600
agttttattt tggtgttgat gtacaagcta accaactgtc aaataagaga ttatgataga      660
ttcgtcattt gctcctttaa tcagctgtcg cgttcccctg ccctataaaa ggagggtatg      720
caccacgatg gttcattacc caataagatt gaaagctcac cactttgttg aaattgacag      780
caaacaaaca aaaaaatgca tttcaccctt tgacatcacc atgcactgcc attaatatgc      840
gccccgttca cacgattcct ctgtagttca gtcggtagaa cggcggactg ttaatccgta      900
tgtcactggt tcgagtccag tcagaggagc caaattcaaa aaagcctgct ttctagcagg      960
ctttttgctt tctaattacc aacgctctta aaacatctgt cttgaaccag aactaatttg     1020
cacaggcatt cccgatcgac gttgcaacgc agcatttg                             1058

<210>  119
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  119
caatgtcgca atttcctgtg                                                   20

<210>  120
<211>  19
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer

<400>  120
caaatgctgc gttgcaacg                                                    19

<210>  121
<211>  4104
<212>  DNA
<213>  Streptococcus  pyogenes

<400>  121
atggataaaa aatacagcat tggtctggat atcggaacca acagcgttgg gtgggcagta       60
ataacagatg aatacaaagt gccgtcaaaa aaatttaagg ttctggggaa tacagatcgc      120
cacagcataa aaaagaatct gattggggca ttgctgtttg attcgggtga gacagctgag      180
gccacgcgtc tgaaacgtac agcaagaaga cgttacacac gtcgtaaaaa tcgtatttgc      240
tacttacagg aaattttttc taacgaaatg gccaaggtag atgatagttt cttccatcgt      300
ctcgaagaat cttttctggt tgaggaagat aaaaaacacg aacgtcaccc tatctttggc      360
aatatcgtgg atgaagtggc ctatcatgaa aaatacccta cgatttatca tcttcgcaag      420
aagttggttg atagtacgga caaagcggat ctgcgtttaa tctatcttgc gttagcgcac      480
atgatcaaat ttcgtggtca tttcttaatt gaaggtgatc tgaatcctga taactctgat      540
gtggacaaat tgtttataca attagtgcaa acctataatc agctgttcga ggaaaacccc      600
attaatgcct ctggagttga tgccaaagcg attttaagcg cgagactttc taagtcccgg      660
cgtctggaga atctgatcgc ccagttacca ggggaaaaga aaaatggtct gtttggtaat      720
ctgattgccc tcagtctggg gcttaccccg aacttcaaat ccaattttga cctggctgag      780
gacgcaaagc tgcagctgag caaagatact tatgatgatg acctcgacaa tctgctcgcc      840
cagattggtg accaatatgc ggatctgttt ctggcagcga agaatctttc ggatgctatc      900
ttgctgtcgg atattctgcg tgttaatacc gaaatcacca aagcgcctct gtctgcaagt      960
atgatcaaga gatacgacga gcaccaccag gacctgactc ttcttaaggc actggtacgc     1020
caacagcttc cggagaaata caaagaaata ttcttcgacc agtccaagaa tggttacgcg     1080
ggctacatcg atggtggtgc atcacaggaa gagttctata aatttattaa accaatcctt     1140
gagaaaatgg atggcacgga agagttactt gttaaactta accgcgaaga cttgcttaga     1200
aagcaacgta cattcgacaa cggctccatc ccacaccaga ttcatttagg tgaacttcac     1260
gccatcttgc gcagacaaga agatttctat cccttcttaa aagacaatcg ggagaaaatc     1320
gagaagatcc tgacgttccg cattccctat tatgtcggtc ccctggcacg tggtaattct     1380
cggtttgcct ggatgacgcg caaaagtgag gaaaccatca ccccttggaa ctttgaagaa     1440
gtcgtggata aaggtgctag cgcgcagtct tttatagaaa gaatgacgaa cttcgataaa     1500
aacttgccca acgaaaaagt cctgcccaag cactctcttt tatatgagta ctttactgtg     1560
tacaacgaac tgactaaagt gaaatacgtt acggaaggta tgcgcaaacc tgcctttctt     1620
agtggcgagc agaaaaaagc aattgtcgat cttctcttta aaacgaatcg caaggtaact     1680
gtaaaacagc tgaaggaaga ttatttcaaa aagatcgaat gctttgattc tgtcgagatc     1740
tcgggtgtcg aagatcgttt caacgcttcc ttagggacct atcatgattt gctgaagata     1800
ataaaagaca aagactttct cgacaatgaa gaaaatgaag atattctgga ggatattgtt     1860
ttgaccttga ccttattcga agatagagag atgatcgagg agcgcttaaa aacctatgcc     1920
cacctgtttg atgacaaagt catgaagcaa ttaaagcgcc gcagatatac ggggtggggc     1980
cgcttgagcc gcaagttgat taacggtatt agagacaagc agagcggaaa aactatcctg     2040
gatttcctca aatctgacgg atttgcgaac cgcaatttta tgcagcttat acatgatgat     2100
tcgcttacat tcaaagagga tattcagaag gctcaggtgt ctgggcaagg tgattcactc     2160
cacgaacata tagcaaattt ggccggctct cctgcgatta agaaggggat cctgcaaaca     2220
gttaaagttg tggatgaact tgtaaaagta atgggccgcc acaagccgga gaatatcgtg     2280
atagaaatgg cgcgcgagaa tcaaacgaca caaaaaggtc aaaagaactc aagagagaga     2340
atgaagcgca ttgaggaggg gataaaggaa cttggatctc aaattctgaa agaacatcca     2400
gttgaaaaca ctcagctgca aaatgaaaaa ttgtacctgt actacctgca gaatggaaga     2460
gacatgtacg tggatcagga attggatatc aatagactct cggactatga cgtagatcac     2520
attgtccctc agagcttcct caaggatgat tctatagata ataaagtact tacgagatcg     2580
gacaaaaatc gcggtaaatc ggataacgtc ccatcggagg aagtcgttaa aaagatgaaa     2640
aactattggc gtcaactgct gaacgccaag ctgatcacac agcgtaagtt tgataatctg     2700
actaaagccg aacgcggtgg tcttagtgaa ctcgataaag caggatttat aaaacggcag     2760
ttagtagaaa cgcgccaaat tacgaaacac gtggctcaga tcctcgattc tagaatgaat     2820
acaaagtacg atgaaaacga taaactgatc cgtgaagtaa aagtcattac cttaaaatct     2880
aaacttgtgt ccgatttccg caaagatttt cagttttaca aggtccggga aatcaataac     2940
tatcaccatg cacatgatgc atatttaaat gcggttgtag gcacggccct tattaagaaa     3000
taccctaaac tcgaaagtga gtttgtttat ggggattata aagtgtatga cgttcgcaaa     3060
atgatcgcga aatcagaaca ggaaatcggt aaggctaccg ctaaatactt tttttattcc     3120
aacattatga atttttttaa gaccgaaata actctcgcga atggtgaaat ccgtaaacgg     3180
cctcttatag aaaccaatgg tgaaacggga gaaatcgttt gggataaagg tcgtgacttt     3240
gccaccgttc gtaaagtcct ctcaatgccg caagttaaca ttgtcaagaa gacggaagtt     3300
caaacagggg gattctccaa agaatctatc ctgccgaagc gtaacagtga taaacttatt     3360
gccagaaaaa aagattggga tccaaaaaaa tacggaggct ttgattcccc taccgtcgcg     3420
tatagtgtgc tggtggttgc taaagtcgag aaagggaaaa gcaagaaatt gaaatcagtt     3480
aaagaactgc tgggtattac aattatggaa agatcgtcct ttgagaaaaa tccgatcgac     3540
tttttagagg ccaaggggta taaggaagtg aaaaaagatc tcatcatcaa attaccgaag     3600
tatagtcttt ttgagctgga aaacggcaga aaaagaatgc tggcctccgc gggcgagtta     3660
cagaagggaa atgagctggc gctgccttcc aaatatgtta attttctgta ccttgccagt     3720
cattatgaga aactgaaggg cagccccgaa gataacgaac agaaacaatt attcgtggaa     3780
cagcataagc actatttaga tgaaattata gagcaaatta gtgaattttc taagcgcgtt     3840
atcctcgcgg atgctaattt agacaaagta ctgtcagctt ataataaaca tcgggataag     3900
ccgattagag aacaggccga aaatatcatt catttgttta ccttaaccaa ccttggagca     3960
ccagctgcct tcaaatattt cgataccaca attgatcgta aacggtatac aagtacaaaa     4020
gaagtcttgg acgcaaccct cattcatcaa tctattactg gattatatga gacacgcatt     4080
gatctttcac agctgggcgg agac                                            4104

<210>  122
<211>  4104
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155H DNA

<400>  122
atggataaaa aatacagcat tggtctggat atcggaacca acagcgttgg gtgggcagta       60
ataacagatg aatacaaagt gccgtcaaaa aaatttaagg ttctggggaa tacagatcgc      120
cacagcataa aaaagaatct gattggggca ttgctgtttg attcgggtga gacagctgag      180
gccacgcgtc tgaaacgtac agcaagaaga cgttacacac gtcgtaaaaa tcgtatttgc      240
tacttacagg aaattttttc taacgaaatg gccaaggtag atgatagttt cttccatcgt      300
ctcgaagaat cttttctggt tgaggaagat aaaaaacacg aacgtcaccc tatctttggc      360
aatatcgtgg atgaagtggc ctatcatgaa aaatacccta cgatttatca tcttcgcaag      420
aagttggttg atagtacgga caaagcggat ctgcgtttaa tccatcttgc gttagcgcac      480
atgatcaaat ttcgtggtca tttcttaatt gaaggtgatc tgaatcctga taactctgat      540
gtggacaaat tgtttataca attagtgcaa acctataatc agctgttcga ggaaaacccc      600
attaatgcct ctggagttga tgccaaagcg attttaagcg cgagactttc taagtcccgg      660
cgtctggaga atctgatcgc ccagttacca ggggaaaaga aaaatggtct gtttggtaat      720
ctgattgccc tcagtctggg gcttaccccg aacttcaaat ccaattttga cctggctgag      780
gacgcaaagc tgcagctgag caaagatact tatgatgatg acctcgacaa tctgctcgcc      840
cagattggtg accaatatgc ggatctgttt ctggcagcga agaatctttc ggatgctatc      900
ttgctgtcgg atattctgcg tgttaatacc gaaatcacca aagcgcctct gtctgcaagt      960
atgatcaaga gatacgacga gcaccaccag gacctgactc ttcttaaggc actggtacgc     1020
caacagcttc cggagaaata caaagaaata ttcttcgacc agtccaagaa tggttacgcg     1080
ggctacatcg atggtggtgc atcacaggaa gagttctata aatttattaa accaatcctt     1140
gagaaaatgg atggcacgga agagttactt gttaaactta accgcgaaga cttgcttaga     1200
aagcaacgta cattcgacaa cggctccatc ccacaccaga ttcatttagg tgaacttcac     1260
gccatcttgc gcagacaaga agatttctat cccttcttaa aagacaatcg ggagaaaatc     1320
gagaagatcc tgacgttccg cattccctat tatgtcggtc ccctggcacg tggtaattct     1380
cggtttgcct ggatgacgcg caaaagtgag gaaaccatca ccccttggaa ctttgaagaa     1440
gtcgtggata aaggtgctag cgcgcagtct tttatagaaa gaatgacgaa cttcgataaa     1500
aacttgccca acgaaaaagt cctgcccaag cactctcttt tatatgagta ctttactgtg     1560
tacaacgaac tgactaaagt gaaatacgtt acggaaggta tgcgcaaacc tgcctttctt     1620
agtggcgagc agaaaaaagc aattgtcgat cttctcttta aaacgaatcg caaggtaact     1680
gtaaaacagc tgaaggaaga ttatttcaaa aagatcgaat gctttgattc tgtcgagatc     1740
tcgggtgtcg aagatcgttt caacgcttcc ttagggacct atcatgattt gctgaagata     1800
ataaaagaca aagactttct cgacaatgaa gaaaatgaag atattctgga ggatattgtt     1860
ttgaccttga ccttattcga agatagagag atgatcgagg agcgcttaaa aacctatgcc     1920
cacctgtttg atgacaaagt catgaagcaa ttaaagcgcc gcagatatac ggggtggggc     1980
cgcttgagcc gcaagttgat taacggtatt agagacaagc agagcggaaa aactatcctg     2040
gatttcctca aatctgacgg atttgcgaac cgcaatttta tgcagcttat acatgatgat     2100
tcgcttacat tcaaagagga tattcagaag gctcaggtgt ctgggcaagg tgattcactc     2160
cacgaacata tagcaaattt ggccggctct cctgcgatta agaaggggat cctgcaaaca     2220
gttaaagttg tggatgaact tgtaaaagta atgggccgcc acaagccgga gaatatcgtg     2280
atagaaatgg cgcgcgagaa tcaaacgaca caaaaaggtc aaaagaactc aagagagaga     2340
atgaagcgca ttgaggaggg gataaaggaa cttggatctc aaattctgaa agaacatcca     2400
gttgaaaaca ctcagctgca aaatgaaaaa ttgtacctgt actacctgca gaatggaaga     2460
gacatgtacg tggatcagga attggatatc aatagactct cggactatga cgtagatcac     2520
attgtccctc agagcttcct caaggatgat tctatagata ataaagtact tacgagatcg     2580
gacaaaaatc gcggtaaatc ggataacgtc ccatcggagg aagtcgttaa aaagatgaaa     2640
aactattggc gtcaactgct gaacgccaag ctgatcacac agcgtaagtt tgataatctg     2700
actaaagccg aacgcggtgg tcttagtgaa ctcgataaag caggatttat aaaacggcag     2760
ttagtagaaa cgcgccaaat tacgaaacac gtggctcaga tcctcgattc tagaatgaat     2820
acaaagtacg atgaaaacga taaactgatc cgtgaagtaa aagtcattac cttaaaatct     2880
aaacttgtgt ccgatttccg caaagatttt cagttttaca aggtccggga aatcaataac     2940
tatcaccatg cacatgatgc atatttaaat gcggttgtag gcacggccct tattaagaaa     3000
taccctaaac tcgaaagtga gtttgtttat ggggattata aagtgtatga cgttcgcaaa     3060
atgatcgcga aatcagaaca ggaaatcggt aaggctaccg ctaaatactt tttttattcc     3120
aacattatga atttttttaa gaccgaaata actctcgcga atggtgaaat ccgtaaacgg     3180
cctcttatag aaaccaatgg tgaaacggga gaaatcgttt gggataaagg tcgtgacttt     3240
gccaccgttc gtaaagtcct ctcaatgccg caagttaaca ttgtcaagaa gacggaagtt     3300
caaacagggg gattctccaa agaatctatc ctgccgaagc gtaacagtga taaacttatt     3360
gccagaaaaa aagattggga tccaaaaaaa tacggaggct ttgattcccc taccgtcgcg     3420
tatagtgtgc tggtggttgc taaagtcgag aaagggaaaa gcaagaaatt gaaatcagtt     3480
aaagaactgc tgggtattac aattatggaa agatcgtcct ttgagaaaaa tccgatcgac     3540
tttttagagg ccaaggggta taaggaagtg aaaaaagatc tcatcatcaa attaccgaag     3600
tatagtcttt ttgagctgga aaacggcaga aaaagaatgc tggcctccgc gggcgagtta     3660
cagaagggaa atgagctggc gctgccttcc aaatatgtta attttctgta ccttgccagt     3720
cattatgaga aactgaaggg cagccccgaa gataacgaac agaaacaatt attcgtggaa     3780
cagcataagc actatttaga tgaaattata gagcaaatta gtgaattttc taagcgcgtt     3840
atcctcgcgg atgctaattt agacaaagta ctgtcagctt ataataaaca tcgggataag     3900
ccgattagag aacaggccga aaatatcatt catttgttta ccttaaccaa ccttggagca     3960
ccagctgcct tcaaatattt cgataccaca attgatcgta aacggtatac aagtacaaaa     4020
gaagtcttgg acgcaaccct cattcatcaa tctattactg gattatatga gacacgcatt     4080
gatctttcac agctgggcgg agac                                            4104

<210>  123
<211>  1368
<212>  PRT
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155N variant

<400>  123
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Asn Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             
Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             
Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             
Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             
Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             
Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             
Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             
Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             
Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             
Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             
Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             
Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             
Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             
Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             
Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             
Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             
Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             
Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             
His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             
Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             
Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             
Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             
Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             
Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             
Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             

<210>  124
<211>  4104
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155N

<400>  124
atggataaaa aatacagcat tggtctggat atcggaacca acagcgttgg gtgggcagta       60
ataacagatg aatacaaagt gccgtcaaaa aaatttaagg ttctggggaa tacagatcgc      120
cacagcataa aaaagaatct gattggggca ttgctgtttg attcgggtga gacagctgag      180
gccacgcgtc tgaaacgtac agcaagaaga cgttacacac gtcgtaaaaa tcgtatttgc      240
tacttacagg aaattttttc taacgaaatg gccaaggtag atgatagttt cttccatcgt      300
ctcgaagaat cttttctggt tgaggaagat aaaaaacacg aacgtcaccc tatctttggc      360
aatatcgtgg atgaagtggc ctatcatgaa aaatacccta cgatttatca tcttcgcaag      420
aagttggttg atagtacgga caaagcggat ctgcgtttaa tcaatcttgc gttagcgcac      480
atgatcaaat ttcgtggtca tttcttaatt gaaggtgatc tgaatcctga taactctgat      540
gtggacaaat tgtttataca attagtgcaa acctataatc agctgttcga ggaaaacccc      600
attaatgcct ctggagttga tgccaaagcg attttaagcg cgagactttc taagtcccgg      660
cgtctggaga atctgatcgc ccagttacca ggggaaaaga aaaatggtct gtttggtaat      720
ctgattgccc tcagtctggg gcttaccccg aacttcaaat ccaattttga cctggctgag      780
gacgcaaagc tgcagctgag caaagatact tatgatgatg acctcgacaa tctgctcgcc      840
cagattggtg accaatatgc ggatctgttt ctggcagcga agaatctttc ggatgctatc      900
ttgctgtcgg atattctgcg tgttaatacc gaaatcacca aagcgcctct gtctgcaagt      960
atgatcaaga gatacgacga gcaccaccag gacctgactc ttcttaaggc actggtacgc     1020
caacagcttc cggagaaata caaagaaata ttcttcgacc agtccaagaa tggttacgcg     1080
ggctacatcg atggtggtgc atcacaggaa gagttctata aatttattaa accaatcctt     1140
gagaaaatgg atggcacgga agagttactt gttaaactta accgcgaaga cttgcttaga     1200
aagcaacgta cattcgacaa cggctccatc ccacaccaga ttcatttagg tgaacttcac     1260
gccatcttgc gcagacaaga agatttctat cccttcttaa aagacaatcg ggagaaaatc     1320
gagaagatcc tgacgttccg cattccctat tatgtcggtc ccctggcacg tggtaattct     1380
cggtttgcct ggatgacgcg caaaagtgag gaaaccatca ccccttggaa ctttgaagaa     1440
gtcgtggata aaggtgctag cgcgcagtct tttatagaaa gaatgacgaa cttcgataaa     1500
aacttgccca acgaaaaagt cctgcccaag cactctcttt tatatgagta ctttactgtg     1560
tacaacgaac tgactaaagt gaaatacgtt acggaaggta tgcgcaaacc tgcctttctt     1620
agtggcgagc agaaaaaagc aattgtcgat cttctcttta aaacgaatcg caaggtaact     1680
gtaaaacagc tgaaggaaga ttatttcaaa aagatcgaat gctttgattc tgtcgagatc     1740
tcgggtgtcg aagatcgttt caacgcttcc ttagggacct atcatgattt gctgaagata     1800
ataaaagaca aagactttct cgacaatgaa gaaaatgaag atattctgga ggatattgtt     1860
ttgaccttga ccttattcga agatagagag atgatcgagg agcgcttaaa aacctatgcc     1920
cacctgtttg atgacaaagt catgaagcaa ttaaagcgcc gcagatatac ggggtggggc     1980
cgcttgagcc gcaagttgat taacggtatt agagacaagc agagcggaaa aactatcctg     2040
gatttcctca aatctgacgg atttgcgaac cgcaatttta tgcagcttat acatgatgat     2100
tcgcttacat tcaaagagga tattcagaag gctcaggtgt ctgggcaagg tgattcactc     2160
cacgaacata tagcaaattt ggccggctct cctgcgatta agaaggggat cctgcaaaca     2220
gttaaagttg tggatgaact tgtaaaagta atgggccgcc acaagccgga gaatatcgtg     2280
atagaaatgg cgcgcgagaa tcaaacgaca caaaaaggtc aaaagaactc aagagagaga     2340
atgaagcgca ttgaggaggg gataaaggaa cttggatctc aaattctgaa agaacatcca     2400
gttgaaaaca ctcagctgca aaatgaaaaa ttgtacctgt actacctgca gaatggaaga     2460
gacatgtacg tggatcagga attggatatc aatagactct cggactatga cgtagatcac     2520
attgtccctc agagcttcct caaggatgat tctatagata ataaagtact tacgagatcg     2580
gacaaaaatc gcggtaaatc ggataacgtc ccatcggagg aagtcgttaa aaagatgaaa     2640
aactattggc gtcaactgct gaacgccaag ctgatcacac agcgtaagtt tgataatctg     2700
actaaagccg aacgcggtgg tcttagtgaa ctcgataaag caggatttat aaaacggcag     2760
ttagtagaaa cgcgccaaat tacgaaacac gtggctcaga tcctcgattc tagaatgaat     2820
acaaagtacg atgaaaacga taaactgatc cgtgaagtaa aagtcattac cttaaaatct     2880
aaacttgtgt ccgatttccg caaagatttt cagttttaca aggtccggga aatcaataac     2940
tatcaccatg cacatgatgc atatttaaat gcggttgtag gcacggccct tattaagaaa     3000
taccctaaac tcgaaagtga gtttgtttat ggggattata aagtgtatga cgttcgcaaa     3060
atgatcgcga aatcagaaca ggaaatcggt aaggctaccg ctaaatactt tttttattcc     3120
aacattatga atttttttaa gaccgaaata actctcgcga atggtgaaat ccgtaaacgg     3180
cctcttatag aaaccaatgg tgaaacggga gaaatcgttt gggataaagg tcgtgacttt     3240
gccaccgttc gtaaagtcct ctcaatgccg caagttaaca ttgtcaagaa gacggaagtt     3300
caaacagggg gattctccaa agaatctatc ctgccgaagc gtaacagtga taaacttatt     3360
gccagaaaaa aagattggga tccaaaaaaa tacggaggct ttgattcccc taccgtcgcg     3420
tatagtgtgc tggtggttgc taaagtcgag aaagggaaaa gcaagaaatt gaaatcagtt     3480
aaagaactgc tgggtattac aattatggaa agatcgtcct ttgagaaaaa tccgatcgac     3540
tttttagagg ccaaggggta taaggaagtg aaaaaagatc tcatcatcaa attaccgaag     3600
tatagtcttt ttgagctgga aaacggcaga aaaagaatgc tggcctccgc gggcgagtta     3660
cagaagggaa atgagctggc gctgccttcc aaatatgtta attttctgta ccttgccagt     3720
cattatgaga aactgaaggg cagccccgaa gataacgaac agaaacaatt attcgtggaa     3780
cagcataagc actatttaga tgaaattata gagcaaatta gtgaattttc taagcgcgtt     3840
atcctcgcgg atgctaattt agacaaagta ctgtcagctt ataataaaca tcgggataag     3900
ccgattagag aacaggccga aaatatcatt catttgttta ccttaaccaa ccttggagca     3960
ccagctgcct tcaaatattt cgataccaca attgatcgta aacggtatac aagtacaaaa     4020
gaagtcttgg acgcaaccct cattcatcaa tctattactg gattatatga gacacgcatt     4080
gatctttcac agctgggcgg agac                                            4104

<210>  125
<211>  1368
<212>  PRT
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155E

<400>  125
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Glu Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             
Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             
Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             
Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             
Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             
Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             
Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             
Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             
Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             
Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             
Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             
Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             
Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             
Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             
Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             
Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             
Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             
Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             
His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             
Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             
Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             
Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             
Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             
Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             
Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             

<210>  126
<211>  4104
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155E

<400>  126
atggataaaa aatacagcat tggtctggat atcggaacca acagcgttgg gtgggcagta       60
ataacagatg aatacaaagt gccgtcaaaa aaatttaagg ttctggggaa tacagatcgc      120
cacagcataa aaaagaatct gattggggca ttgctgtttg attcgggtga gacagctgag      180
gccacgcgtc tgaaacgtac agcaagaaga cgttacacac gtcgtaaaaa tcgtatttgc      240
tacttacagg aaattttttc taacgaaatg gccaaggtag atgatagttt cttccatcgt      300
ctcgaagaat cttttctggt tgaggaagat aaaaaacacg aacgtcaccc tatctttggc      360
aatatcgtgg atgaagtggc ctatcatgaa aaatacccta cgatttatca tcttcgcaag      420
aagttggttg atagtacgga caaagcggat ctgcgtttaa tcgagcttgc gttagcgcac      480
atgatcaaat ttcgtggtca tttcttaatt gaaggtgatc tgaatcctga taactctgat      540
gtggacaaat tgtttataca attagtgcaa acctataatc agctgttcga ggaaaacccc      600
attaatgcct ctggagttga tgccaaagcg attttaagcg cgagactttc taagtcccgg      660
cgtctggaga atctgatcgc ccagttacca ggggaaaaga aaaatggtct gtttggtaat      720
ctgattgccc tcagtctggg gcttaccccg aacttcaaat ccaattttga cctggctgag      780
gacgcaaagc tgcagctgag caaagatact tatgatgatg acctcgacaa tctgctcgcc      840
cagattggtg accaatatgc ggatctgttt ctggcagcga agaatctttc ggatgctatc      900
ttgctgtcgg atattctgcg tgttaatacc gaaatcacca aagcgcctct gtctgcaagt      960
atgatcaaga gatacgacga gcaccaccag gacctgactc ttcttaaggc actggtacgc     1020
caacagcttc cggagaaata caaagaaata ttcttcgacc agtccaagaa tggttacgcg     1080
ggctacatcg atggtggtgc atcacaggaa gagttctata aatttattaa accaatcctt     1140
gagaaaatgg atggcacgga agagttactt gttaaactta accgcgaaga cttgcttaga     1200
aagcaacgta cattcgacaa cggctccatc ccacaccaga ttcatttagg tgaacttcac     1260
gccatcttgc gcagacaaga agatttctat cccttcttaa aagacaatcg ggagaaaatc     1320
gagaagatcc tgacgttccg cattccctat tatgtcggtc ccctggcacg tggtaattct     1380
cggtttgcct ggatgacgcg caaaagtgag gaaaccatca ccccttggaa ctttgaagaa     1440
gtcgtggata aaggtgctag cgcgcagtct tttatagaaa gaatgacgaa cttcgataaa     1500
aacttgccca acgaaaaagt cctgcccaag cactctcttt tatatgagta ctttactgtg     1560
tacaacgaac tgactaaagt gaaatacgtt acggaaggta tgcgcaaacc tgcctttctt     1620
agtggcgagc agaaaaaagc aattgtcgat cttctcttta aaacgaatcg caaggtaact     1680
gtaaaacagc tgaaggaaga ttatttcaaa aagatcgaat gctttgattc tgtcgagatc     1740
tcgggtgtcg aagatcgttt caacgcttcc ttagggacct atcatgattt gctgaagata     1800
ataaaagaca aagactttct cgacaatgaa gaaaatgaag atattctgga ggatattgtt     1860
ttgaccttga ccttattcga agatagagag atgatcgagg agcgcttaaa aacctatgcc     1920
cacctgtttg atgacaaagt catgaagcaa ttaaagcgcc gcagatatac ggggtggggc     1980
cgcttgagcc gcaagttgat taacggtatt agagacaagc agagcggaaa aactatcctg     2040
gatttcctca aatctgacgg atttgcgaac cgcaatttta tgcagcttat acatgatgat     2100
tcgcttacat tcaaagagga tattcagaag gctcaggtgt ctgggcaagg tgattcactc     2160
cacgaacata tagcaaattt ggccggctct cctgcgatta agaaggggat cctgcaaaca     2220
gttaaagttg tggatgaact tgtaaaagta atgggccgcc acaagccgga gaatatcgtg     2280
atagaaatgg cgcgcgagaa tcaaacgaca caaaaaggtc aaaagaactc aagagagaga     2340
atgaagcgca ttgaggaggg gataaaggaa cttggatctc aaattctgaa agaacatcca     2400
gttgaaaaca ctcagctgca aaatgaaaaa ttgtacctgt actacctgca gaatggaaga     2460
gacatgtacg tggatcagga attggatatc aatagactct cggactatga cgtagatcac     2520
attgtccctc agagcttcct caaggatgat tctatagata ataaagtact tacgagatcg     2580
gacaaaaatc gcggtaaatc ggataacgtc ccatcggagg aagtcgttaa aaagatgaaa     2640
aactattggc gtcaactgct gaacgccaag ctgatcacac agcgtaagtt tgataatctg     2700
actaaagccg aacgcggtgg tcttagtgaa ctcgataaag caggatttat aaaacggcag     2760
ttagtagaaa cgcgccaaat tacgaaacac gtggctcaga tcctcgattc tagaatgaat     2820
acaaagtacg atgaaaacga taaactgatc cgtgaagtaa aagtcattac cttaaaatct     2880
aaacttgtgt ccgatttccg caaagatttt cagttttaca aggtccggga aatcaataac     2940
tatcaccatg cacatgatgc atatttaaat gcggttgtag gcacggccct tattaagaaa     3000
taccctaaac tcgaaagtga gtttgtttat ggggattata aagtgtatga cgttcgcaaa     3060
atgatcgcga aatcagaaca ggaaatcggt aaggctaccg ctaaatactt tttttattcc     3120
aacattatga atttttttaa gaccgaaata actctcgcga atggtgaaat ccgtaaacgg     3180
cctcttatag aaaccaatgg tgaaacggga gaaatcgttt gggataaagg tcgtgacttt     3240
gccaccgttc gtaaagtcct ctcaatgccg caagttaaca ttgtcaagaa gacggaagtt     3300
caaacagggg gattctccaa agaatctatc ctgccgaagc gtaacagtga taaacttatt     3360
gccagaaaaa aagattggga tccaaaaaaa tacggaggct ttgattcccc taccgtcgcg     3420
tatagtgtgc tggtggttgc taaagtcgag aaagggaaaa gcaagaaatt gaaatcagtt     3480
aaagaactgc tgggtattac aattatggaa agatcgtcct ttgagaaaaa tccgatcgac     3540
tttttagagg ccaaggggta taaggaagtg aaaaaagatc tcatcatcaa attaccgaag     3600
tatagtcttt ttgagctgga aaacggcaga aaaagaatgc tggcctccgc gggcgagtta     3660
cagaagggaa atgagctggc gctgccttcc aaatatgtta attttctgta ccttgccagt     3720
cattatgaga aactgaaggg cagccccgaa gataacgaac agaaacaatt attcgtggaa     3780
cagcataagc actatttaga tgaaattata gagcaaatta gtgaattttc taagcgcgtt     3840
atcctcgcgg atgctaattt agacaaagta ctgtcagctt ataataaaca tcgggataag     3900
ccgattagag aacaggccga aaatatcatt catttgttta ccttaaccaa ccttggagca     3960
ccagctgcct tcaaatattt cgataccaca attgatcgta aacggtatac aagtacaaaa     4020
gaagtcttgg acgcaaccct cattcatcaa tctattactg gattatatga gacacgcatt     4080
gatctttcac agctgggcgg agac                                            4104

<210>  127
<211>  1368
<212>  PRT
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155F

<400>  127
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Phe Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             
Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             
Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             
Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             
Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             
Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             
Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             
Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             
Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             
Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             
Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             
Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             
Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             
Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             
Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             
Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             
Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             
Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             
His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             
Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             
Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             
Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             
Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             
Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             
Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             

<210>  128
<211>  4104
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 Y155F

<400>  128
atggataaaa aatacagcat tggtctggat atcggaacca acagcgttgg gtgggcagta       60
ataacagatg aatacaaagt gccgtcaaaa aaatttaagg ttctggggaa tacagatcgc      120
cacagcataa aaaagaatct gattggggca ttgctgtttg attcgggtga gacagctgag      180
gccacgcgtc tgaaacgtac agcaagaaga cgttacacac gtcgtaaaaa tcgtatttgc      240
tacttacagg aaattttttc taacgaaatg gccaaggtag atgatagttt cttccatcgt      300
ctcgaagaat cttttctggt tgaggaagat aaaaaacacg aacgtcaccc tatctttggc      360
aatatcgtgg atgaagtggc ctatcatgaa aaatacccta cgatttatca tcttcgcaag      420
aagttggttg atagtacgga caaagcggat ctgcgtttaa tctttcttgc gttagcgcac      480
atgatcaaat ttcgtggtca tttcttaatt gaaggtgatc tgaatcctga taactctgat      540
gtggacaaat tgtttataca attagtgcaa acctataatc agctgttcga ggaaaacccc      600
attaatgcct ctggagttga tgccaaagcg attttaagcg cgagactttc taagtcccgg      660
cgtctggaga atctgatcgc ccagttacca ggggaaaaga aaaatggtct gtttggtaat      720
ctgattgccc tcagtctggg gcttaccccg aacttcaaat ccaattttga cctggctgag      780
gacgcaaagc tgcagctgag caaagatact tatgatgatg acctcgacaa tctgctcgcc      840
cagattggtg accaatatgc ggatctgttt ctggcagcga agaatctttc ggatgctatc      900
ttgctgtcgg atattctgcg tgttaatacc gaaatcacca aagcgcctct gtctgcaagt      960
atgatcaaga gatacgacga gcaccaccag gacctgactc ttcttaaggc actggtacgc     1020
caacagcttc cggagaaata caaagaaata ttcttcgacc agtccaagaa tggttacgcg     1080
ggctacatcg atggtggtgc atcacaggaa gagttctata aatttattaa accaatcctt     1140
gagaaaatgg atggcacgga agagttactt gttaaactta accgcgaaga cttgcttaga     1200
aagcaacgta cattcgacaa cggctccatc ccacaccaga ttcatttagg tgaacttcac     1260
gccatcttgc gcagacaaga agatttctat cccttcttaa aagacaatcg ggagaaaatc     1320
gagaagatcc tgacgttccg cattccctat tatgtcggtc ccctggcacg tggtaattct     1380
cggtttgcct ggatgacgcg caaaagtgag gaaaccatca ccccttggaa ctttgaagaa     1440
gtcgtggata aaggtgctag cgcgcagtct tttatagaaa gaatgacgaa cttcgataaa     1500
aacttgccca acgaaaaagt cctgcccaag cactctcttt tatatgagta ctttactgtg     1560
tacaacgaac tgactaaagt gaaatacgtt acggaaggta tgcgcaaacc tgcctttctt     1620
agtggcgagc agaaaaaagc aattgtcgat cttctcttta aaacgaatcg caaggtaact     1680
gtaaaacagc tgaaggaaga ttatttcaaa aagatcgaat gctttgattc tgtcgagatc     1740
tcgggtgtcg aagatcgttt caacgcttcc ttagggacct atcatgattt gctgaagata     1800
ataaaagaca aagactttct cgacaatgaa gaaaatgaag atattctgga ggatattgtt     1860
ttgaccttga ccttattcga agatagagag atgatcgagg agcgcttaaa aacctatgcc     1920
cacctgtttg atgacaaagt catgaagcaa ttaaagcgcc gcagatatac ggggtggggc     1980
cgcttgagcc gcaagttgat taacggtatt agagacaagc agagcggaaa aactatcctg     2040
gatttcctca aatctgacgg atttgcgaac cgcaatttta tgcagcttat acatgatgat     2100
tcgcttacat tcaaagagga tattcagaag gctcaggtgt ctgggcaagg tgattcactc     2160
cacgaacata tagcaaattt ggccggctct cctgcgatta agaaggggat cctgcaaaca     2220
gttaaagttg tggatgaact tgtaaaagta atgggccgcc acaagccgga gaatatcgtg     2280
atagaaatgg cgcgcgagaa tcaaacgaca caaaaaggtc aaaagaactc aagagagaga     2340
atgaagcgca ttgaggaggg gataaaggaa cttggatctc aaattctgaa agaacatcca     2400
gttgaaaaca ctcagctgca aaatgaaaaa ttgtacctgt actacctgca gaatggaaga     2460
gacatgtacg tggatcagga attggatatc aatagactct cggactatga cgtagatcac     2520
attgtccctc agagcttcct caaggatgat tctatagata ataaagtact tacgagatcg     2580
gacaaaaatc gcggtaaatc ggataacgtc ccatcggagg aagtcgttaa aaagatgaaa     2640
aactattggc gtcaactgct gaacgccaag ctgatcacac agcgtaagtt tgataatctg     2700
actaaagccg aacgcggtgg tcttagtgaa ctcgataaag caggatttat aaaacggcag     2760
ttagtagaaa cgcgccaaat tacgaaacac gtggctcaga tcctcgattc tagaatgaat     2820
acaaagtacg atgaaaacga taaactgatc cgtgaagtaa aagtcattac cttaaaatct     2880
aaacttgtgt ccgatttccg caaagatttt cagttttaca aggtccggga aatcaataac     2940
tatcaccatg cacatgatgc atatttaaat gcggttgtag gcacggccct tattaagaaa     3000
taccctaaac tcgaaagtga gtttgtttat ggggattata aagtgtatga cgttcgcaaa     3060
atgatcgcga aatcagaaca ggaaatcggt aaggctaccg ctaaatactt tttttattcc     3120
aacattatga atttttttaa gaccgaaata actctcgcga atggtgaaat ccgtaaacgg     3180
cctcttatag aaaccaatgg tgaaacggga gaaatcgttt gggataaagg tcgtgacttt     3240
gccaccgttc gtaaagtcct ctcaatgccg caagttaaca ttgtcaagaa gacggaagtt     3300
caaacagggg gattctccaa agaatctatc ctgccgaagc gtaacagtga taaacttatt     3360
gccagaaaaa aagattggga tccaaaaaaa tacggaggct ttgattcccc taccgtcgcg     3420
tatagtgtgc tggtggttgc taaagtcgag aaagggaaaa gcaagaaatt gaaatcagtt     3480
aaagaactgc tgggtattac aattatggaa agatcgtcct ttgagaaaaa tccgatcgac     3540
tttttagagg ccaaggggta taaggaagtg aaaaaagatc tcatcatcaa attaccgaag     3600
tatagtcttt ttgagctgga aaacggcaga aaaagaatgc tggcctccgc gggcgagtta     3660
cagaagggaa atgagctggc gctgccttcc aaatatgtta attttctgta ccttgccagt     3720
cattatgaga aactgaaggg cagccccgaa gataacgaac agaaacaatt attcgtggaa     3780
cagcataagc actatttaga tgaaattata gagcaaatta gtgaattttc taagcgcgtt     3840
atcctcgcgg atgctaattt agacaaagta ctgtcagctt ataataaaca tcgggataag     3900
ccgattagag aacaggccga aaatatcatt catttgttta ccttaaccaa ccttggagca     3960
ccagctgcct tcaaatattt cgataccaca attgatcgta aacggtatac aagtacaaaa     4020
gaagtcttgg acgcaaccct cattcatcaa tctattactg gattatatga gacacgcatt     4080
gatctttcac agctgggcgg agac                                            4104

<210>  129
<211>  1368
<212>  PRT
<213>  Artificial sequence

<220>
<223>  synthesized Cas9 F86A F98A variant

<400>  129
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Ala Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Ala Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile His Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             
Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             
Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             
Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             
Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             
Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             
Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             
Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             
Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             
Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             
Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             
Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             
Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             
Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             
Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             
Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             
Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             
Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             
His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             
Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             
Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             
Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             
Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             
Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             
Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             

<210>  130
<211>  498
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthesized F86A F98A synthetic fragment

<400>  130
cacgtcgtaa aaatcgtatt tgctacttac aggaaattgc gtctaacgaa atggccaagg       60
tagatgatag tgcgttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac      120
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc      180
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt      240
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg      300
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata      360
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa      420
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa      480
agaaaaatgg tctgtttg                                                    498

<210>  131
<211>  9266
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid pRF801 backbone for F86A-F98A

<400>  131
aaagaaaaat ggtctgtttg gtaatctgat tgccctcagt ctggggctta ccccgaactt       60
caaatccaat tttgacctgg ctgaggacgc aaagctgcag ctgagcaaag atacttatga      120
tgatgacctc gacaatctgc tcgcccagat tggtgaccaa tatgcggatc tgtttctggc      180
agcgaagaat ctttcggatg ctatcttgct gtcggatatt ctgcgtgtta ataccgaaat      240
caccaaagcg cctctgtctg caagtatgat caagagatac gacgagcacc accaggacct      300
gactcttctt aaggcactgg tacgccaaca gcttccggag aaatacaaag aaatattctt      360
cgaccagtcc aagaatggtt acgcgggcta catcgatggt ggtgcatcac aggaagagtt      420
ctataaattt attaaaccaa tccttgagaa aatggatggc acggaagagt tacttgttaa      480
acttaaccgc gaagacttgc ttagaaagca acgtacattc gacaacggct ccatcccaca      540
ccagattcat ttaggtgaac ttcacgccat cttgcgcaga caagaagatt tctatccctt      600
cttaaaagac aatcgggaga aaatcgagaa gatcctgacg ttccgcattc cctattatgt      660
cggtcccctg gcacgtggta attctcggtt tgcctggatg acgcgcaaaa gtgaggaaac      720
catcacccct tggaactttg aagaagtcgt ggataaaggt gctagcgcgc agtcttttat      780
agaaagaatg acgaacttcg ataaaaactt gcccaacgaa aaagtcctgc ccaagcactc      840
tcttttatat gagtacttta ctgtgtacaa cgaactgact aaagtgaaat acgttacgga      900
aggtatgcgc aaacctgcct ttcttagtgg cgagcagaaa aaagcaattg tcgatcttct      960
ctttaaaacg aatcgcaagg taactgtaaa acagctgaag gaagattatt tcaaaaagat     1020
cgaatgcttt gattctgtcg agatctcggg tgtcgaagat cgtttcaacg cttccttagg     1080
gacctatcat gatttgctga agataataaa agacaaagac tttctcgaca atgaagaaaa     1140
tgaagatatt ctggaggata ttgttttgac cttgacctta ttcgaagata gagagatgat     1200
cgaggagcgc ttaaaaacct atgcccacct gtttgatgac aaagtcatga agcaattaaa     1260
gcgccgcaga tatacggggt ggggccgctt gagccgcaag ttgattaacg gtattagaga     1320
caagcagagc ggaaaaacta tcctggattt cctcaaatct gacggatttg cgaaccgcaa     1380
ttttatgcag cttatacatg atgattcgct tacattcaaa gaggatattc agaaggctca     1440
ggtgtctggg caaggtgatt cactccacga acatatagca aatttggccg gctctcctgc     1500
gattaagaag gggatcctgc aaacagttaa agttgtggat gaacttgtaa aagtaatggg     1560
ccgccacaag ccggagaata tcgtgataga aatggcgcgc gagaatcaaa cgacacaaaa     1620
aggtcaaaag aactcaagag agagaatgaa gcgcattgag gaggggataa aggaacttgg     1680
atctcaaatt ctgaaagaac atccagttga aaacactcag ctgcaaaatg aaaaattgta     1740
cctgtactac ctgcagaatg gaagagacat gtacgtggat caggaattgg atatcaatag     1800
actctcggac tatgacgtag atcacattgt ccctcagagc ttcctcaagg atgattctat     1860
agataataaa gtacttacga gatcggacaa aaatcgcggt aaatcggata acgtcccatc     1920
ggaggaagtc gttaaaaaga tgaaaaacta ttggcgtcaa ctgctgaacg ccaagctgat     1980
cacacagcgt aagtttgata atctgactaa agccgaacgc ggtggtctta gtgaactcga     2040
taaagcagga tttataaaac ggcagttagt agaaacgcgc caaattacga aacacgtggc     2100
tcagatcctc gattctagaa tgaatacaaa gtacgatgaa aacgataaac tgatccgtga     2160
agtaaaagtc attaccttaa aatctaaact tgtgtccgat ttccgcaaag attttcagtt     2220
ttacaaggtc cgggaaatca ataactatca ccatgcacat gatgcatatt taaatgcggt     2280
tgtaggcacg gcccttatta agaaataccc taaactcgaa agtgagtttg tttatgggga     2340
ttataaagtg tatgacgttc gcaaaatgat cgcgaaatca gaacaggaaa tcggtaaggc     2400
taccgctaaa tacttttttt attccaacat tatgaatttt tttaagaccg aaataactct     2460
cgcgaatggt gaaatccgta aacggcctct tatagaaacc aatggtgaaa cgggagaaat     2520
cgtttgggat aaaggtcgtg actttgccac cgttcgtaaa gtcctctcaa tgccgcaagt     2580
taacattgtc aagaagacgg aagttcaaac agggggattc tccaaagaat ctatcctgcc     2640
gaagcgtaac agtgataaac ttattgccag aaaaaaagat tgggatccaa aaaaatacgg     2700
aggctttgat tcccctaccg tcgcgtatag tgtgctggtg gttgctaaag tcgagaaagg     2760
gaaaagcaag aaattgaaat cagttaaaga actgctgggt attacaatta tggaaagatc     2820
gtcctttgag aaaaatccga tcgacttttt agaggccaag gggtataagg aagtgaaaaa     2880
agatctcatc atcaaattac cgaagtatag tctttttgag ctggaaaacg gcagaaaaag     2940
aatgctggcc tccgcgggcg agttacagaa gggaaatgag ctggcgctgc cttccaaata     3000
tgttaatttt ctgtaccttg ccagtcatta tgagaaactg aagggcagcc ccgaagataa     3060
cgaacagaaa caattattcg tggaacagca taagcactat ttagatgaaa ttatagagca     3120
aattagtgaa ttttctaagc gcgttatcct cgcggatgct aatttagaca aagtactgtc     3180
agcttataat aaacatcggg ataagccgat tagagaacag gccgaaaata tcattcattt     3240
gtttacctta accaaccttg gagcaccagc tgccttcaaa tatttcgata ccacaattga     3300
tcgtaaacgg tatacaagta caaaagaagt cttggacgca accctcattc atcaatctat     3360
tactggatta tatgagacac gcattgatct ttcacagctg ggcggagaca agaagaaaaa     3420
actgaaactg caccatcatc accatcatca tcaccatcat tgataactcg agaaagctta     3480
cataaaaaac cggccttggc cccgccggtt ttttattatt tttcttcctc cgcatgttca     3540
atccgctcca taatcgacgg atggctccct ctgaaaattt taacgagaaa cggcgggttg     3600
acccggctca gtcccgtaac ggccaagtcc tgaaacgtct caatcgccgc ttcccggttt     3660
ccggtcagct caatgccgta acggtcggcg gcgttttcct gataccggga gacggcattc     3720
gtaatcgggt gaagtggtca agacctcact aggcacctta aaaatagcgc accctgaaga     3780
agatttattt gaggtagccc ttgcctacct agcttccaag aaagatatcc taacagcaca     3840
agagcggaaa gatgttttgt tctacatcca gaacaacctc tgctaaaatt cctgaaaaat     3900
tttgcaaaaa gttgttgact ttatctacaa ggtgtggcat aatgtgtgga ctcgacttcg     3960
aatacatcca gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac     4020
ttgaaaaagt ggcaccgagt cggtgcgact cctgttgata gatccagtaa tgacctcaga     4080
actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa     4140
tgtcgacctc gagagttacg ctagggataa cagggtaata taggagctcc agtcggctta     4200
aaccagtttt cgctggtgcg aaaaaagagt gtcttgtgac acctaaattc aaaatctatc     4260
ggtcagattt ataccgattt gattttatat attcttgaat aacatacgcc gagttatcac     4320
ataaaagcgg gaaccaatca taaaatttaa acttcattgc ataatccatt aaactcttaa     4380
attctacgat tccttgttca tcaataaact caatcatttc tttaattaat ttatatctat     4440
ctgttgttgt tttctttaat aattcattaa catctacacc gccataaact atcatatctt     4500
ctttttgata tttaaattta ttaggatcgt ccatgtgaag catatatctc acaagacctt     4560
tcacacttcc tgcaatctgc ggaatagtcg cattcaattc ttctgttaat tatttttatc     4620
tgttcataag atttattacc ctcatacatc actagaatat gataatgctc ttttttcatc     4680
ctaccttctg tatcagtatc cctatcatgt aatggagaca ctacaaattg aatgtgtaac     4740
tcttttaaat actctaacca ctcggctttt gctgattctg gatataaaac aaatgtccaa     4800
ttacgtcctc ttgaattttt cttgttttca gtttctttta ttacattttc gctcatgata     4860
taataacggt gctaatacac ttaacaaaat ttagtcatag ataggcagca tgccagtgct     4920
gtctatcttt ttttgtttaa aatgcaccgt attcctcctt tgcatatttt tttattagaa     4980
taccggttgc atctgatttg ctaatattat atttttcttt gattctattt aatatctcat     5040
tttcttctgt tgtaagtctt aaagtaacag caactttttt ctcttctttt ctatctacaa     5100
ctatcactgt acctcccaac atctgttttt ttcactttaa cataaaaaac aaccttttaa     5160
cattaaaaac ccaatattta tttatttgtt tggacaatgg acactggaca cctagggggg     5220
aggtcgtagt acccccctat gttttctccc ctaaataacc ccaaaaatct aagaaaaaaa     5280
gacctcaaaa aggtctttaa ttaacatctc aaatttcgca tttattccaa tttccttttt     5340
gcgtgtgatg cgagctcatc ggctccgtcg atactatgtt atacgccaac tttcaaaaca     5400
actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac agtgaattgg     5460
agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga aaagaggaag     5520
gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga tcgaaaaata     5580
ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc tggtgggaga     5640
aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca cctatgatgt     5700
ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc caaaggtcct     5760
gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg atggcgtcct     5820
ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc tgtatgcgga     5880
gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga atagcttaga     5940
cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg tggattgcga     6000
aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt ttttaaagac     6060
ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca gcaacatctt     6120
tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca gggcggacaa     6180
gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg aagaacagta     6240
tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa taaaatatta     6300
tattttactg gatgaattgt tttagtgact gcagtgagat ctggtaatga ctctctagct     6360
tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt     6420
gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctctagcta agcagaaggc     6480
catcctgacg gatggccttt ttgcgtttct acaaactctt gttaactcta gagctgcctg     6540
ccgcgtttcg gtgatgaaga tcttcccgat gattaattaa ttcagaacgc tcggttgccg     6600
ccgggcgttt tttatgaagc ttcgttgctg gcgtttttcc ataggctccg cccccctgac     6660
gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga     6720
taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt     6780
accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc     6840
tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc     6900
cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta     6960
agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat     7020
gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca     7080
gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct     7140
tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt     7200
acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct     7260
cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc     7320
acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa     7380
acttggtctg acaaatggtt ctttcccctg tcctaaacaa aaaacccgct ttattgaaaa     7440
agcggggctg ttttacagac aggtcaaata aacgtttgaa aatgttcatt tcaaaacgcg     7500
cggaacctcc atcttctccc atccagacta tactgtcggc ttcggaatcg caccgaatcc     7560
tgcccataaa aaggctcgcg ggcttagagc gcttgctcat caccgccggt agggaatttc     7620
accctgcccc gaagattgat cttatttatt tttaatactg atattattat aaattaattg     7680
tgaaaaaatg tacaggtgca aagcttattg cgctgttttg ggacatcctg cacgatattt     7740
cggtaaactc actttttccg catactaaaa accgcacatt cacagttatt tcatttttaa     7800
ttttcgtctt tccgcgtgaa actcattgac actctttatg gaatatggta aattatcaga     7860
tatttatgac gcttatttag gaggaaatct tacacagaag ctgcggaacc tgaaaagaat     7920
tcctttcagg ttccgttttt tttaggaatt ctccctgatc tcaagcatct ggcggggata     7980
aatccgctct cctttcaaat cgttccattc tttgaggcgc tgtacagtta cgcccatttt     8040
ttcggcgata tgatgaagcg tatccccttt ccgcactaca tatgtaccgg tcttcgattc     8100
atcgtcatga aggcggagtg tttggccggc cttgagattt gaatgtttca acccgtttat     8160
tctcatgatc tcctcgatgg atataccgct atccttgctg attctccaga gcgtgtcccc     8220
tttttgaacg gtcaccgcac cgctcattgt cccggcgttt tgataaacgt ggatagaatt     8280
ttgccggaac gcctcctcac gaagcaccgt cagcggattg attgcatatc ttttatcttc     8340
agtccatgaa ccgtgatgca tttcaaaatg caggtgggtt ccggtcgata ttcgaattcc     8400
tccattttct tctgctatca aaataacaga ctcgtgattt tccaaacgag ctttcaaaaa     8460
agcctctgcc ccttgcaaat cggatgcctg tctataaaat tcccgatatt ggttaaacag     8520
cggcgcaatg gcggccgcat ctgatgtctt tgcttggcga atgttcatct tatttcttcc     8580
tccctctcaa taattttttc attctatccc ttttctgtaa agtttatttt tcagaatact     8640
tttatcatca tgctttgaaa aaatatcacg ataatatcca ttgttctcac ggaagcacac     8700
gcaggtcatt tgaacgaatt ttttcgacag gaatttgccg ggactcagga gcatttaacc     8760
taaaaaagca tgacatttca gcataatgaa catttactca tgtctatttt cgttcttttc     8820
tgtatgaaaa tagttatttc gagtctctac ggaaatagcg agagatgata tacctaaata     8880
gagataaaat catctcaaaa aaatgggtct actaaaatat tattccatct attacaataa     8940
attcacagaa tagtctttta agtaagtcta ctctgaattt ttttaaaagg agagggtaac     9000
tagtggcccc aaaaaagaaa cgcaaggtta tggataaaaa atacagcatt ggtctggata     9060
tcggaaccaa cagcgttggg tgggcagtaa taacagatga atacaaagtg ccgtcaaaaa     9120
aatttaaggt tctggggaat acagatcgcc acagcataaa aaagaatctg attggggcat     9180
tgctgtttga ttcgggtgag acagctgagg ccacgcgtct gaaacgtaca gcaagaagac     9240
gttacacacg tcgtaaaaat cgtatt                                          9266

<210>  132
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer pRF801 backbone forward

<400>  132
aaagaaaaat ggtctgtttg                                                   20

<210>  133
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer pRF801 backbone reverse

<400>  133
aatacgattt ttacgacgtg                                                   20

<210>  134
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer F86A F98A synthetic forward

<400>  134
cacgtcgtaa aaatcgtatt                                                   20

<210>  135
<211>  20
<212>  DNA
<213>  Artificial sequence

<220>
<223>  primer F86A F98A synthetic reverse

<400>  135
caaacagacc atttttcttt                                                   20

<210>  136
<211>  9724
<212>  DNA
<213>  Artificial sequence

<220>
<223>  plasmid  pRF866

<400>  136
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt       60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg      120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca      180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca      240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa      300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca      360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga      420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag      480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag      540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa      600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta      660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg      720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt      780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac      840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca      900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct      960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt     1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt     1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa     1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat     1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg     1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt     1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca     1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa     1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg     1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc     1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt     1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg     1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg     1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata     1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg     1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga     1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg     1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt     2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc     2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat     2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg     2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg     2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa     2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa     2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta     2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga     2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt     2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc     2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt     2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct     2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt     2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc     2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat     2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag     3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga     3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg     3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt     3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac     3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc     3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt     3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc     3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc     3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg     3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag     3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg     3660
tctgacaaat ggttctttcc cctgtcctaa acaaaaaacc cgctttattg aaaaagcggg     3720
gctgttttac agacaggtca aataaacgtt tgaaaatgtt catttcaaaa cgcgcggaac     3780
ctccatcttc tcccatccag actatactgt cggcttcgga atcgcaccga atcctgccca     3840
taaaaaggct cgcgggctta gagcgcttgc tcatcaccgc cggtagggaa tttcaccctg     3900
ccccgaagat tgatcttatt tatttttaat actgatatta ttataaatta attgtgaaaa     3960
aatgtacagg tgcaaagctt attgcgctgt tttgggacat cctgcacgat atttcggtaa     4020
actcactttt tccgcatact aaaaaccgca cattcacagt tatttcattt ttaattttcg     4080
tctttccgcg tgaaactcat tgacactctt tatggaatat ggtaaattat cagatattta     4140
tgacgcttat ttaggaggaa atcttacaca gaagctgcgg aacctgaaaa gaattccttt     4200
caggttccgt tttttttagg aattctccct gatctcaagc atctggcggg gataaatccg     4260
ctctcctttc aaatcgttcc attctttgag gcgctgtaca gttacgccca ttttttcggc     4320
gatatgatga agcgtatccc ctttccgcac tacatatgta ccggtcttcg attcatcgtc     4380
atgaaggcgg agtgtttggc cggccttgag atttgaatgt ttcaacccgt ttattctcat     4440
gatctcctcg atggatatac cgctatcctt gctgattctc cagagcgtgt cccctttttg     4500
aacggtcacc gcaccgctca ttgtcccggc gttttgataa acgtggatag aattttgccg     4560
gaacgcctcc tcacgaagca ccgtcagcgg attgattgca tatcttttat cttcagtcca     4620
tgaaccgtga tgcatttcaa aatgcaggtg ggttccggtc gatattcgaa ttcctccatt     4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc     4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc     4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc     4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc     4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt     4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa     5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg     5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata     5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac     5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg     5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa     5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta     5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt     5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca     5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaattgc gtctaacgaa atggccaagg     5580
tagatgatag tgcgttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac     5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc     5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt     5760
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg     5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata     5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa     5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa     6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca     6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg     6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag     6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca     6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga     6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg     6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct     6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac     6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc     6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct     6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg     6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca     6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag     6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc     6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag     6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct     6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg     7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga     7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg     7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg     7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc     7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca     7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt     7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg     7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga     7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc     7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag     7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat     7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc     7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac     7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag     7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg     7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca     7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata     8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc     8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag     8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt     8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg     8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt     8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta     8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg     8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg     8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta     8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga     8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag     8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga     8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt     8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag     8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa     8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg     9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg     9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa     9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag     9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt     9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc     9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta     9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac     9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca     9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat     9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac     9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc     9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt     9720
aatc                                                                  9724

<210>  137
<211>  5042
<212>  DNA
<213>  Artificial sequence

<220>
<223>   Bacillus F86A-F98A expression cassette

<400>  137
attcctccat tttcttctgc tatcaaaata acagactcgt gattttccaa acgagctttc       60
aaaaaagcct ctgccccttg caaatcggat gcctgtctat aaaattcccg atattggtta      120
aacagcggcg caatggcggc cgcatctgat gtctttgctt ggcgaatgtt catcttattt      180
cttcctccct ctcaataatt ttttcattct atcccttttc tgtaaagttt atttttcaga      240
atacttttat catcatgctt tgaaaaaata tcacgataat atccattgtt ctcacggaag      300
cacacgcagg tcatttgaac gaattttttc gacaggaatt tgccgggact caggagcatt      360
taacctaaaa aagcatgaca tttcagcata atgaacattt actcatgtct attttcgttc      420
ttttctgtat gaaaatagtt atttcgagtc tctacggaaa tagcgagaga tgatatacct      480
aaatagagat aaaatcatct caaaaaaatg ggtctactaa aatattattc catctattac      540
aataaattca cagaatagtc ttttaagtaa gtctactctg aattttttta aaaggagagg      600
gtaactagtg gccccaaaaa agaaacgcaa ggttatggat aaaaaataca gcattggtct      660
ggatatcgga accaacagcg ttgggtgggc agtaataaca gatgaataca aagtgccgtc      720
aaaaaaattt aaggttctgg ggaatacaga tcgccacagc ataaaaaaga atctgattgg      780
ggcattgctg tttgattcgg gtgagacagc tgaggccacg cgtctgaaac gtacagcaag      840
aagacgttac acacgtcgta aaaatcgtat ttgctactta caggaaattg cgtctaacga      900
aatggccaag gtagatgata gtgcgttcca tcgtctcgaa gaatcttttc tggttgagga      960
agataaaaaa cacgaacgtc accctatctt tggcaatatc gtggatgaag tggcctatca     1020
tgaaaaatac cctacgattt atcatcttcg caagaagttg gttgatagta cggacaaagc     1080
ggatctgcgt ttaatccatc ttgcgttagc gcacatgatc aaatttcgtg gtcatttctt     1140
aattgaaggt gatctgaatc ctgataactc tgatgtggac aaattgttta tacaattagt     1200
gcaaacctat aatcagctgt tcgaggaaaa ccccattaat gcctctggag ttgatgccaa     1260
agcgatttta agcgcgagac tttctaagtc ccggcgtctg gagaatctga tcgcccagtt     1320
accaggggaa aagaaaaatg gtctgtttgg taatctgatt gccctcagtc tggggcttac     1380
cccgaacttc aaatccaatt ttgacctggc tgaggacgca aagctgcagc tgagcaaaga     1440
tacttatgat gatgacctcg acaatctgct cgcccagatt ggtgaccaat atgcggatct     1500
gtttctggca gcgaagaatc tttcggatgc tatcttgctg tcggatattc tgcgtgttaa     1560
taccgaaatc accaaagcgc ctctgtctgc aagtatgatc aagagatacg acgagcacca     1620
ccaggacctg actcttctta aggcactggt acgccaacag cttccggaga aatacaaaga     1680
aatattcttc gaccagtcca agaatggtta cgcgggctac atcgatggtg gtgcatcaca     1740
ggaagagttc tataaattta ttaaaccaat ccttgagaaa atggatggca cggaagagtt     1800
acttgttaaa cttaaccgcg aagacttgct tagaaagcaa cgtacattcg acaacggctc     1860
catcccacac cagattcatt taggtgaact tcacgccatc ttgcgcagac aagaagattt     1920
ctatcccttc ttaaaagaca atcgggagaa aatcgagaag atcctgacgt tccgcattcc     1980
ctattatgtc ggtcccctgg cacgtggtaa ttctcggttt gcctggatga cgcgcaaaag     2040
tgaggaaacc atcacccctt ggaactttga agaagtcgtg gataaaggtg ctagcgcgca     2100
gtcttttata gaaagaatga cgaacttcga taaaaacttg cccaacgaaa aagtcctgcc     2160
caagcactct cttttatatg agtactttac tgtgtacaac gaactgacta aagtgaaata     2220
cgttacggaa ggtatgcgca aacctgcctt tcttagtggc gagcagaaaa aagcaattgt     2280
cgatcttctc tttaaaacga atcgcaaggt aactgtaaaa cagctgaagg aagattattt     2340
caaaaagatc gaatgctttg attctgtcga gatctcgggt gtcgaagatc gtttcaacgc     2400
ttccttaggg acctatcatg atttgctgaa gataataaaa gacaaagact ttctcgacaa     2460
tgaagaaaat gaagatattc tggaggatat tgttttgacc ttgaccttat tcgaagatag     2520
agagatgatc gaggagcgct taaaaaccta tgcccacctg tttgatgaca aagtcatgaa     2580
gcaattaaag cgccgcagat atacggggtg gggccgcttg agccgcaagt tgattaacgg     2640
tattagagac aagcagagcg gaaaaactat cctggatttc ctcaaatctg acggatttgc     2700
gaaccgcaat tttatgcagc ttatacatga tgattcgctt acattcaaag aggatattca     2760
gaaggctcag gtgtctgggc aaggtgattc actccacgaa catatagcaa atttggccgg     2820
ctctcctgcg attaagaagg ggatcctgca aacagttaaa gttgtggatg aacttgtaaa     2880
agtaatgggc cgccacaagc cggagaatat cgtgatagaa atggcgcgcg agaatcaaac     2940
gacacaaaaa ggtcaaaaga actcaagaga gagaatgaag cgcattgagg aggggataaa     3000
ggaacttgga tctcaaattc tgaaagaaca tccagttgaa aacactcagc tgcaaaatga     3060
aaaattgtac ctgtactacc tgcagaatgg aagagacatg tacgtggatc aggaattgga     3120
tatcaataga ctctcggact atgacgtaga tcacattgtc cctcagagct tcctcaagga     3180
tgattctata gataataaag tacttacgag atcggacaaa aatcgcggta aatcggataa     3240
cgtcccatcg gaggaagtcg ttaaaaagat gaaaaactat tggcgtcaac tgctgaacgc     3300
caagctgatc acacagcgta agtttgataa tctgactaaa gccgaacgcg gtggtcttag     3360
tgaactcgat aaagcaggat ttataaaacg gcagttagta gaaacgcgcc aaattacgaa     3420
acacgtggct cagatcctcg attctagaat gaatacaaag tacgatgaaa acgataaact     3480
gatccgtgaa gtaaaagtca ttaccttaaa atctaaactt gtgtccgatt tccgcaaaga     3540
ttttcagttt tacaaggtcc gggaaatcaa taactatcac catgcacatg atgcatattt     3600
aaatgcggtt gtaggcacgg cccttattaa gaaataccct aaactcgaaa gtgagtttgt     3660
ttatggggat tataaagtgt atgacgttcg caaaatgatc gcgaaatcag aacaggaaat     3720
cggtaaggct accgctaaat acttttttta ttccaacatt atgaattttt ttaagaccga     3780
aataactctc gcgaatggtg aaatccgtaa acggcctctt atagaaacca atggtgaaac     3840
gggagaaatc gtttgggata aaggtcgtga ctttgccacc gttcgtaaag tcctctcaat     3900
gccgcaagtt aacattgtca agaagacgga agttcaaaca gggggattct ccaaagaatc     3960
tatcctgccg aagcgtaaca gtgataaact tattgccaga aaaaaagatt gggatccaaa     4020
aaaatacgga ggctttgatt cccctaccgt cgcgtatagt gtgctggtgg ttgctaaagt     4080
cgagaaaggg aaaagcaaga aattgaaatc agttaaagaa ctgctgggta ttacaattat     4140
ggaaagatcg tcctttgaga aaaatccgat cgacttttta gaggccaagg ggtataagga     4200
agtgaaaaaa gatctcatca tcaaattacc gaagtatagt ctttttgagc tggaaaacgg     4260
cagaaaaaga atgctggcct ccgcgggcga gttacagaag ggaaatgagc tggcgctgcc     4320
ttccaaatat gttaattttc tgtaccttgc cagtcattat gagaaactga agggcagccc     4380
cgaagataac gaacagaaac aattattcgt ggaacagcat aagcactatt tagatgaaat     4440
tatagagcaa attagtgaat tttctaagcg cgttatcctc gcggatgcta atttagacaa     4500
agtactgtca gcttataata aacatcggga taagccgatt agagaacagg ccgaaaatat     4560
cattcatttg tttaccttaa ccaaccttgg agcaccagct gccttcaaat atttcgatac     4620
cacaattgat cgtaaacggt atacaagtac aaaagaagtc ttggacgcaa ccctcattca     4680
tcaatctatt actggattat atgagacacg cattgatctt tcacagctgg gcggagacaa     4740
gaagaaaaaa ctgaaactgc accatcatca ccatcatcat caccatcatt gataaacata     4800
aaaaaccggc cttggccccg ccggtttttt attatttttc ttcctccgca tgttcaatcc     4860
gctccataat cgacggatgg ctccctctga aaattttaac gagaaacggc gggttgaccc     4920
ggctcagtcc cgtaacggcc aagtcctgaa acgtctcaat cgccgcttcc cggtttccgg     4980
tcagctcaat gccgtaacgg tcggcggcgt tttcctgata ccgggagacg gcattcgtaa     5040
tc                                                                    5042

<210>  138
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  138
aaagaaatat atagagagat actcttatca atgatggtga tgatgatggt gatg             54

<210>  139
<211>  58
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  139
acacgtattt atttgtccaa ttaccatggc cccaaaaaag aaacgcaagg ttatggat         58

<210>  140
<211>  767
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized RNR2p promoter

<400>  140
gccaatctaa agcgttgtct tcctcgttcc tcgcgcacgc gattttgcgt tctgcatagg       60
aagccgaagt cgaacaagaa gcaggcaaag tttagagcac tgcccctccg cactcaaaaa      120
agaaaaaact aggaggaaaa taaaattctc aaccacacaa acacataaac acatacaaat      180
acaaatacaa gcttatttac ttgacatcgc gcgatcttcc actattcagc gccgtccgcc      240
ctctctcgtg ttttttgttt acgcgacaac tatgcgaaat ccggagcaac gggcaaccgt      300
ttggggaaag accacaccca cgcgcgatcg ccatggcaac gaggtcgcac acgccccaca      360
cccagacctc cctgcgagcg ggcatgggta caatgtcccc gttgccacag agaccacttc      420
gtagcacagc gcagagcgta gcgtgttgtt gctgctgaca aaagaaaatt tttcttagca      480
aagcaaagga ggggaagcac gggcagatag caccgtacca tacccttgga aactcgaaat      540
gaacgaagca ggaaatgaga gaatgagagt tttgtaggta tatatagcgg tagtgtttgc      600
gcgttaccat catcttctgg atctatctat tgttcttttc ctcatcactt tccccttttt      660
cgctcttctt cttgtctttt atttctttct tttttttaat tgttccctcg attggctatc      720
taccaaagaa tccaaactta atacacgtat ttatttgtcc aattacc                    767

<210>  141
<211>  1346
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized 2-micron replication origin 1

<400>  141
aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt       60
caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt      120
taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa      180
tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc      240
tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc      300
gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg      360
cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg      420
tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta      480
gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat      540
gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag      600
aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca      660
ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa      720
gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg      780
agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga      840
tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc      900
cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc      960
tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt     1020
ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac     1080
gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg     1140
cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct     1200
agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta     1260
ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct     1320
atcatttcct ttgatattgg atcata                                          1346

<210>  142
<211>  1705
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  KanMX expression cassette

<400>  142
ctagggattc ataaccattt tctcaatcga attacacaga acacaccgta caaacctctc       60
tatcataact acttaatagt cacacacgta ctcgtctaaa tacacatcat cgtcctacaa      120
gttcatcaaa gtgttggaca gacaactata ccagcatgga tctcttgtat cggttctttt      180
ctcccgctct ctcgcaataa caatgaacac tgggtcaatc atagcctaca caggtgaaca      240
gagtagcgtt tatacagggt ttatacggtg attcctacgg caaaaatttt tcatttctaa      300
aaaaaaaaag aaaaattttt ctttccaacg ctagaaggaa aagaaaaatc taattaaatt      360
gatttggtga ttttctgaga gttccctttt tcatatatcg aattttgaat ataaaaggag      420
atcgaaaaaa tttttctatt caatctgttt tctggtttta tttgatagtt tttttgtgta      480
ttattattat ggattagtac tggtttatat gggtttttct gtataacttc tttttatttt      540
agtttgttta atcttatttt gagttacatt atagttccct aactgcaaga gaagtaacat      600
taaaactcga gatgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca      660
tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga      720
caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag      780
gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta      840
tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca      900
ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa      960
atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt     1020
gtccttttaa cagcgatcgc gtatttcgtc tggctcaggc gcaatcacga atgaataacg     1080
gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct     1140
ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt     1200
tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac     1260
gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt     1320
tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga     1380
ataaattgca gtttcatttg atgctcgatg agtttttcta agtttaactt gatactacta     1440
gatttttctc ttcatttata aaatttttgg ttataattga agctttagaa gtatgaaaaa     1500
atcctttttt ttcattcttt gcaaccaaaa taagaagctt cttttattca ttgaaatgat     1560
gaatataaac ctaacaaaag aaaaagactc gaatatcaaa cattaaaaaa aaataaaaga     1620
ggttatctgt tttcccattt agttggagtt tgcattttct aatagataga actctcaatt     1680
aatgtggatt tagtttctct gttcg                                           1705

<210>  143
<211>  299
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized SNR52p promoter

<400>  143
ccctcactaa agggaacaaa agctggagct tctttgaaaa gataatgtat gattatgctt       60
tcactcatat ttatacagaa acttgatgtt ttctttcgag tatatacaag gtgattacat      120
gtacgtttga agtacaactc tagattttgt agtgccctct tgggctagcg ggaaaggtgc      180
gcattttttc acaccctaca atgttctgtt caaaagattt tggtcaaacg ctgtagaagt      240
gaaagttggt gcgcatgttt cggcgttcga aacttctccg cagtgaaaga taaatgatc       299

<210>  144
<211>  11555
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pSE087 plasmid

<400>  144
gatcctctag tttctcggta ctatgcatat gatccaatat caaaggaaat gatagcattg       60
aaggatgaga ctaatccaat tgaggagtgg cagcatatag aacagctaaa gggtagtgct      120
gaaggaagca tacgataccc cgcatggaat gggataatat cacaggaggt actagactac      180
ctttcatcct acataaatag acgcatataa gtacgcattt aagcataaac acgcactatg      240
ccgttcttct catgtatata tatatacagg caacacgcag atataggtgc gacgtgaaca      300
gtgagctgta tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct      360
ttgaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt      420
tgaaaaccaa aagcgctctg aagacgcact ttcaaaaaac caaaaacgca ccggactgta      480
acgagctact aaaatattgc gaataccgct tccacaaaca ttgctcaaaa gtatctcttt      540
gctatatatc tctgtgctat atccctatat aacctaccca tccacctttc gctccttgaa      600
cttgcatcta aactcgacct ctacattttt tatgtttatc tctagtatta ctctttagac      660
aaaaaaattg tagtaagaac tattcataga gtgaatcgaa aacaatacga aaatgtaaac      720
atttcctata cgtagtatat agagacaaaa tagaagaaac cgttcataat tttctgacca      780
atgaagaatc atcaacgcta tcactttctg ttcacaaagt atgcgcaatc cacatcggta      840
tagaatataa tcggggatgc ctttatcttg aaaaaatgca cccgcagctt cgctagtaat      900
cagtaaacgc gggaagtgga gtcaggcttt ttttatggaa gagaaaatag acaccaaagt      960
agccttcttc taaccttaac ggacctacag tgcaaaaagt tatcaagaga ctgcattata     1020
gagcgcacaa aggagaaaaa aagtaatcta agatgctttg ttagaaaaat agcgctctcg     1080
ggatgcattt ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct     1140
cgcgttgcat ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc     1200
tctcgcgttg catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc     1260
gctttcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt     1320
agcgctctcg cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttaacaaag     1380
atatgctatt gaagtgcaag atggaaacgc agaaaatgaa ccggggatgc gacgtgcaag     1440
attacctatg caatagatgc aatagtttcg gcatgccgaa cagagaaact aaatccacat     1500
taattgagag ttctatctat tagaaaatgc aaactccaac taaatgggaa aacagataac     1560
ctcttttatt tttttttaat gtttgatatt cgagtctttt tcttttgtta ggtttatatt     1620
catcatttca atgaataaaa gaagcttctt attttggttg caaagaatga aaaaaaagga     1680
ttttttcata cttctaaagc ttcaattata accaaaaatt ttataaatga agagaaaaat     1740
ctagtagtat caagttaaac ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta     1800
ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa     1860
aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact     1920
cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag     1980
aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc     2040
cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa     2100
ccgttattca ttcgtgattg cgcctgagcc agacgaaata cgcgatcgct gttaaaagga     2160
caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata     2220
ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttgcc ggggatcgca     2280
gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc     2340
ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta     2400
cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt     2460
gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc     2520
atgttggaat ttaatcgcgg cctcgaaacg tgagtctttt ccttacccat ctcgagtttt     2580
aatgttactt ctcttgcagt tagggaacta taatgtaact caaaataaga ttaaacaaac     2640
taaaataaaa agaagttata cagaaaaacc catataaacc agtactaatc cataataata     2700
atacacaaaa aaactatcaa ataaaaccag aaaacagatt gaatagaaaa attttttcga     2760
tctcctttta tattcaaaat tcgatatatg aaaaagggaa ctctcagaaa atcaccaaat     2820
caatttaatt agatttttct tttccttcta gcgttggaaa gaaaaatttt tctttttttt     2880
tttagaaatg aaaaattttt gccgtaggaa tcaccgtata aaccctgtat aaacgctact     2940
ctgttcacct gtgtaggcta tgattgaccc agtgttcatt gttattgcga gagagcggga     3000
gaaaagaacc gatacaagag atccatgctg gtatagttgt ctgtccaaca ctttgatgaa     3060
cttgtaggac gatgatgtgt atttagacga gtacgtgtgt gactattaag tagttatgat     3120
agagaggttt gtacggtgtg ttctgtgtaa ttcgattgag aaaatggtta tgaatcccta     3180
gacccgggaa tctctaagta aatgcatgta tactaaactc acaaattaga gcttcaattt     3240
aattatatca gttattaccc gggttacgcc aagcgcgcaa ttaaccctca ctaaagggaa     3300
caaaagctgg agctcccctc actaaaggga acaaaagctg gagcttcttt gaaaagataa     3360
tgtatgatta tgctttcact catatttata cagaaacttg atgttttctt tcgagtatat     3420
acaaggtgat tacatgtacg tttgaagtac aactctagat tttgtagtgc cctcttgggc     3480
tagcgggaaa ggtgcgcatt ttttcacacc ctacaatgtt ctgttcaaaa gattttggtc     3540
aaacgctgta gaagtgaaag ttggtgcgca tgtttcggcg ttcgaaactt ctccgcagtg     3600
aaagataaat gatcggagac ggatacgttc tctatggagg agttttagag ctagaaatag     3660
caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtggtgc     3720
tttttttctg gccgtctcta agggcccggt acccaattcg ccctatagtg agtcgtatta     3780
cgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc     3840
tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta     3900
tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag     3960
aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg     4020
tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg     4080
tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg     4140
cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga     4200
agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc     4260
tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt     4320
aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact     4380
ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg     4440
cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt     4500
accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt     4560
ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct     4620
ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg     4680
gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt     4740
aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt     4800
gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc     4860
gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg     4920
cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc     4980
gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg     5040
gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca     5100
ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga     5160
tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct     5220
ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg     5280
cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca     5340
accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata     5400
cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct     5460
tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact     5520
cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa     5580
acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc     5640
atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga     5700
tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga     5760
aaagtgccac ctgacgtcgg cgcgccattt aaatagatcc ggtccgctcg agctgtaccg     5820
cagatgggac ctcttatgaa agtagacatc cttaagtgat gtaggggtat gtccgagtag     5880
ttcgttcaac tatttttaaa gacacatcgt tccaggttta tattcatata catatgtatg     5940
tattgtatag gtttttttat tttttatttt ttattttttt tttttttttt ttcaattttt     6000
ttaagctttt cttacttttc cgatgccctt tccactttaa tcaatatacg tatgaaattc     6060
agccgaagtt tccgtttgat aatatattag attcaatata aaaaaaagtt tatatgattc     6120
agtgtatata tatatatata aataagaggt gcgaaagccc acataaagag attgaagaga     6180
ctgcgtaaaa agaaatatat agagagatac tcttaactag acctttcttt tcttctttgg     6240
gtcggctctg gagtcgccac ccaattgaga taagtcaatt ctagtttcgt acaaaccagt     6300
aatagattga tggatcaagg tggcgtccaa gacttccttg gtagaggtgt aacgcttacg     6360
gtcaatagtg gtgtcgaagt acttgaaggc agcaggagca cccaaattgg tcaaagtgaa     6420
caagtgaatg atgttttcag cttgttctct gattggcttg tctctgtgct tgttgtaggc     6480
ggacaaaacc ttgtccaagt tagcgtcagc caaaatgact ctcttggaga actcggagat     6540
ttgttcaatg atttcgtcca agtagtgctt gtgttgttcg acgaatagtt gcttttgttc     6600
gttgtcttct ggactaccct tcaacttttc gtagtgagaa gccaagtata ggaagttaac     6660
gtacttgctt ggcaaggcca attcgttacc cttttgcaat tcaccagcgg aagccaacat     6720
tctctttcta ccgttttcca attcgaacaa tgagtacttt ggcaacttta tgatcaagtc     6780
tttcttgact tccttgtaac ccttagcttc taggaagtcg attgggttct tttcgaaaga     6840
acttctttcc atgatggtaa tacctaacaa ttccttgacg ctcttcaatt tcttagactt     6900
acccttttca accttagcga caaccaaaac gctgtaggca acggtaggag agtcgaagcc     6960
accgtatttc tttgggtccc aatctttctt tctagcgatc aacttatcag agtttctctt     7020
tggtaagata gattccttag agaagccacc agtttggact tcggttttct tgacgatgtt     7080
gacttgtggc atagacaaaa cctttctaac ggtagcgaaa tcacgaccct tgtcccaaac     7140
aatttcacct gtttcaccgt ttgtttcgat caatggtctc tttctgattt caccgttagc     7200
caaggtgatt tcggtcttaa agaagttcat aatgttagag taaaagaagt acttagcagt     7260
ggccttaccg atttcttgtt cagacttagc gatcatcttt ctgacatcgt agaccttgta     7320
atcaccgtag acaaattcag attctagctt tgggtatttc ttgattagag cggtacccac     7380
gacagcgttc aagtaagcat cgtgagcatg gtggtaattg ttgatttctc taaccttgta     7440
gaattggaag tcctttctga agtcagaaac caacttggac ttcaaagtga tgaccttgac     7500
ttctctaatc aacttatcat tttcgtcgta cttggtgttc atacgggaat ctaggatttg     7560
ggcaacgtgc ttggtgattt gtctggtttc gaccaattgt ctcttgatga agccagcctt     7620
atccaattcg gacaatccac ctctttcagc cttagttagg ttatcgaact ttctttgagt     7680
gattagctta gcgtttagca attgtctcca gtagttcttc atctttttca cgacctcttc     7740
ggatggaacg ttgtcagact tacctctgtt cttgtcggat ctggtcaaaa ccttgttgtc     7800
aatagaatcg tccttcaaga aagattgtgg gacgatgtgg tcaacgtcgt agtcggataa     7860
tctgttgata tctaattctt ggtcaacgta catatctcta ccgttttgca agtagtacaa     7920
gtataacttt tcgttttgta gttgggtgtt ttcaacgggg tgttccttca agatttgaga     7980
acccaattcc ttgataccct cttcgattct cttcattctt tctctagagt tcttttgacc     8040
cttttgtgta gtttgatttt ctctagccat ttctatgacg atgttttctg gcttgtgtct     8100
acccataacc ttaactagtt cgtctacaac cttgacagtt tgtagaatac ctttctttat     8160
ggctggggaa ccagccaagt tagcgatgtg ttcgtgcaag gaatcacctt gaccagaaac     8220
ttgagccttt tggatatctt ccttgaaggt caaagaatcg tcgtggatca attgcatgaa     8280
gtttctgttg gcgaaaccat cggacttcaa gaaatccaag atggtcttac cagattgctt     8340
gtctctgata ccgttaatca actttcttga caatctaccc caaccagtgt acctacgtct     8400
tttcaattgc ttcataacct tgtcatcaaa caagtgggcg taggtcttca atctctcttc     8460
gatcatttct ctgtcttcga acaaggttaa agttaggaca atgtcttcca aaatgtcttc     8520
gttctcttcg ttatccaaga agtccttgtc cttaatgatc tttagtaagt cgtggtaggt     8580
acccaaagaa gcgttgaatc tatcttcaac accagagatt tcaacagaat cgaaacattc     8640
gattttcttg aagtagtctt cctttagttg cttaacagtg acctttctgt ttgtcttgaa     8700
taacaagtca acgatagctt tcttttgttc gccagacaag aaggctggct ttctcatacc     8760
ttcagtaacg tacttgacct tggtcaattc gttgtaaaca gtgaagtatt cgtataacaa     8820
ggaatgcttt ggcaagacct tttcgtttgg caagtttttg tcgaagttgg tcattctttc     8880
gatgaaagat tgagcggaag cacccttatc tacgacctct tcgaaattcc aaggggtgat     8940
ggtctcttcg gactttctgg tcatccaagc gaatctggag ttacctctag ctagaggacc     9000
gacgtagtat gggattctaa aagttaggat cttttcaatc ttttctctgt tgtccttcaa     9060
gaatgggtag aagtcttctt gccttctcaa gatagcgtgt agttcaccta agtggatttg     9120
gtgtggaatg gaaccgttgt cgaaggttct ttgctttctt aacaagtctt ctctattcaa     9180
cttaactagc aactcttcgg taccgtccat cttttccaag attggcttga tgaacttgta     9240
gaactcttct tgactagctc caccgtcgat gtaaccagcg taaccgttct tagattgatc     9300
aaagaagatt tccttgtact tttcgggcaa ttgttgtcta accaaagcct tcaatagagt     9360
caagtcttga tggtgttcgt cgtatctctt aatcatggaa gcagacaatg gagccttagt     9420
gatttcagtg ttaactctca aaatgtcaga tagcaaaatg gcatcagaca agttcttggc     9480
agccaagaac aagtcagcgt attggtcacc aatttgggct aacaagttat ctaaatcgtc     9540
atcgtaagtg tccttggaca attgcaactt agcatcttca gccaaatcga agttagactt     9600
gaagtttgga gtcaaaccca aagacaaagc aatcaagtta ccgaacaaac cgtttttctt     9660
ttcacctggt aattgggcaa tcaagttttc caaacgtctg gacttggata atctagcgga     9720
caaaatggcc ttagcgtcaa caccagaggc gttgatagga ttctcttcga acaattggtt     9780
gtaagtttga accaattgga tgaacaactt atcgacgtca gagttgtctg ggttcaagtc     9840
accttcgatt aggaagtgac cacggaactt gatcatgtgg gccaaagcca agtagatcaa     9900
tctcaagtca gccttgtcgg tggagtcaac caatttcttt ctcaagtggt agatagttgg     9960
gtacttttcg tggtaagcga cttcgtcaac gatgttaccg aagattgggt gtctttcgtg    10020
ctttttgtcc tcttcgacca agaaagactc ttccaatctg tgaaagaaag aatcgtcaac    10080
cttagccatt tcattagaga agatttcttg caagtaacag attctgttct tacgtctagt    10140
gtaacgtcta cgggctgttc tcttcaatct agtagcttcg gcagtttcac cagagtcgaa    10200
taacaaagca ccgattaggt ttttcttgat ggagtgtcta tcggtgttac ccaagacctt    10260
gaatttctta gatgggacct tgtattcgtc agtaatgaca gcccaaccaa cagagttggt    10320
accgatgtcc aaaccgatag agtatttctt gtccatggta attggacaaa taaatacgtg    10380
tattaagttt ggattctttg gtagatagcc aatcgaggga acaattaaaa aaaagaaaga    10440
aataaaagac aagaagaaga gcgaaaaagg ggaaagtgat gaggaaaaga acaatagata    10500
gatccagaag atgatggtaa cgcgcaaaca ctaccgctat atatacctac aaaactctca    10560
ttctctcatt tcctgcttcg ttcatttcga gtttccaagg gtatggtacg gtgctatctg    10620
cccgtgcttc ccctcctttg ctttgctaag aaaaattttc ttttgtcagc agcaacaaca    10680
cgctacgctc tgcgctgtgc tacgaagtgg tctctgtggc aacggggaca ttgtacccat    10740
gcccgctcgc agggaggtct gggtgtgggg cgtgtgcgac ctcgttgcca tggcgatcgc    10800
gcgtgggtgt ggtctttccc caaacggttg cccgttgctc cggatttcgc atagttgtcg    10860
cgtaaacaaa aaacacgaga gagggcggac ggcgctgaat agtggaagat cgcgcgatgt    10920
caagtaaata agcttgtatt tgtatttgta tgtgtttatg tgtttgtgtg gttgagaatt    10980
ttattttcct cctagttttt tcttttttga gtgcggaggg gcagtgctct aaactttgcc    11040
tgcttcttgt tcgacttcgg cttcctatgc agaacgcaaa atcgcgtgcg cgaggaacga    11100
ggaagacaac gctttagatt ggcgcggccg cccgcaaatt aaagccttcg agcgtcccaa    11160
aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa    11220
aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata    11280
aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg    11340
gggagggcgt gaatgtaagc gtgacataac taattacatg attaatctag taacaaggct    11400
aagatatcag cctgaaataa agggtggtga agtaataatt aaatcatccg tataaaccta    11460
tacacatata tgaggaaaaa taatacaaaa gtgttttaaa tacagataca tacatgaaca    11520
tatgcacgta tagcgcccaa atgtcggtaa tggga                               11555

<210>  145
<211>  57
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  145
ctccgcagtg aaagataaat gatcgcccaa aatttgttta ctaaaaacac atgtgga          57

<210>  146
<211>  58
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  146
gaattgggta ccgggccctt agagtaaaaa attgtacttg gcggataatg cctttagc         58

<210>  147
<211>  106
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized targeting sgRNA + T(6) terminator

<400>  147
gatacgttct ctatggagga gttttagagc tagaaatagc aagttaaaat aaggctagtc       60
cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct tttttt                     106

<210>  148
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized 50 bp upstream homology arm

<400>  148
cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc                  50

<210>  149
<211>  106
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized URA3 targeting sgRNA + T(6) terminator

<400>  149
tatcttgact gatttttcca gttttagagc tagaaatagc aagttaaaat aaggctagtc       60
cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct tttttt                     106

<210>  150
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized 50 bp downstream homology arm

<400>  150
gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc                  50

<210>  151
<211>  57
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  151
ccgccaagta caatttttta ctctaagggc ccggtaccca attcgcccta tagtgag          57

<210>  152
<211>  57
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  152
catcatcacc atcattgata agagtatctc tctatatatt tctttttacg cagtctc          57

<210>  153
<211>  566
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized 2-micron replication origin 2

<400>  153
gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat       60
caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat      120
actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct      180
acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt      240
cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg      300
gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta      360
cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg      420
gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg      480
tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc      540
tcgtcagggg ggcggagcct atggaa                                           566

<210>  154
<211>  859
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized ampicillin resistant gene

<400>  154
atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct       60
gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca      120
cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc      180
gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc      240
cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg      300
gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta      360
tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc      420
ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt      480
gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg      540
cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct      600
tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc      660
tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct      720
cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac      780
acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc      840
tcactgatta agcattggt                                                   859

<210>  155
<211>  400
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthesized RNR2 terminator

<400>  155
gagtatctct ctatatattt ctttttacgc agtctcttca atctctttat gtgggctttc       60
gcacctctta tttatatata tatatataca ctgaatcata taaacttttt tttatattga      120
atctaatata ttatcaaacg gaaacttcgg ctgaatttca tacgtatatt gattaaagtg      180
gaaagggcat cggaaaagta agaaaagctt aaaaaaattg aaaaaaaaaa aaaaaaaaat      240
aaaaaataaa aaataaaaaa acctatacaa tacatacata tgtatatgaa tataaacctg      300
gaacgatgtg tctttaaaaa tagttgaacg aactactcgg acatacccct acatcactta      360
aggatgtcta ctttcataag aggtcccatc tgcggtacag                            400

<210>  156
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  156
ccttgcgttt cttttttggg gccatggtaa ttggacaaat aaatacgtgt attaag           56

<210>  157
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  157
tgtttttagt aaacaaattt tgggcgatca tttatctttc actgcggaga agtttc           56
