                         SEQUENCE LISTING

<110>  DSM IP Assets B.V.
 
<120>  GUIDE RNA ASSEMBLY VECTOR

<130>  31203-WO-PCT

<160>  119   

<170>  PatentIn version 3.5

<210>  1
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  Preferred termination sequence in yeast

<400>  1
tttttttgtt ttttatgtct                                                   20


<210>  2
<211>  7240
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence to be integrated in the integration locus
       on chromosome 14 of S. cerevisiae, comprising the Cas9 expression
       cassette and the natMX marker and including the 5'- and 3'- 
       flanking regions

<400>  2
ttgagcggac gccgaaacgt tgtttttttt ttgctttagt ccagataata accttttata       60

attttctttt tagggaggaa gaccggtcta agctcttaga ggttctcgca tacccaagta      120

aaagctaaga ccgaagcaaa cacgcaggat aattttccgg tttactctca ctgcagcttt      180

agtatagatg cagatctggt attgagagag gggggccgga acgaaaagat tcaggggaga      240

gaaaggcccg ggcgtcgggg aacaaaacca cttttgcaca ccttaatttt ttagctactc      300

tgtttctccg ggcttgtatg gcacataaat aaaattctgc tactttccat tatctggtca      360

tcacttaccg atgtggagag ccttgccctt gaaaaataag ataagacctc cgattagggt      420

atttgaaatt aaccgctgta gaatgaaata aggacgcttg tttgacattt tttagttgct      480

ttctctatct accaccattt gcatagcttc aaaatgtttc tactcctttt ttactcttcc      540

agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact      600

aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa      660

agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat      720

cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat gacctcccat      780

tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt      840

ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag      900

ttttctagaa ctagtggatc ccccgggaaa aatggacaag aagtactcca ttgggctcga      960

tatcggcaca aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa     1020

aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc     1080

cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg     1140

cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat     1200

ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga     1260

taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga     1320

aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga     1380

cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat     1440

cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca     1500

gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc     1560

aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc     1620

tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc     1680

caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac     1740

ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt     1800

tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac     1860

ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca     1920

agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat     1980

tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga     2040

ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct     2100

ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat     2160

cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta     2220

cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta     2280

ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga     2340

agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc     2400

cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa     2460

acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt     2520

cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga     2580

cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa     2640

aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc     2700

cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga     2760

ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga     2820

gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca     2880

gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat     2940

ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa     3000

ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa     3060

agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag     3120

cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt     3180

aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac     3240

ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga     3300

actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa     3360

gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat     3420

caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga     3480

ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt     3540

cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa     3600

actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga     3660

gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca     3720

cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat     3780

tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt     3840

tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa     3900

tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta     3960

cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg     4020

caaggccacc gctaagtact tcttttacag caatattatg aattttttca agaccgagat     4080

tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg     4140

agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc     4200

gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat     4260

cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa     4320

atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga     4380

gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga     4440

gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt     4500

caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg     4560

gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc     4620

taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga     4680

agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat     4740

cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt     4800

gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat     4860

ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac     4920

catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca     4980

gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg gagacagcag     5040

ggctgacccc aagaagaaga ggaaggtgtg atctcttctc gagtcatgta attagttatg     5100

tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga     5160

caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta     5220

tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata     5280

ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cgggcgtacg     5340

ctgcaggtcg acgaattcta ccgttcgtat aatgtatgct atacgaagtt atagatctgt     5400

ttagcttgcc ttgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct     5460

ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta     5520

gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg     5580

aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga     5640

cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc     5700

actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat     5760

cacatccgaa cataaacaac catgggtacc actcttgacg acacggctta ccggtaccgc     5820

accagtgtcc cgggggacgc cgaggccatc gaggcactgg atgggtcctt caccaccgac     5880

accgtcttcc gcgtcaccgc caccggggac ggcttcaccc tgcgggaggt gccggtggac     5940

ccgcccctga ccaaggtgtt ccccgacgac gaatcggacg acgaatcgga cgacggggag     6000

gacggcgacc cggactcccg gacgttcgtc gcgtacgggg acgacggcga cctggcgggc     6060

ttcgtggtcg tctcgtactc cggctggaac cgccggctga ccgtcgagga catcgaggtc     6120

gccccggagc accgggggca cggggtcggg cgcgcgttga tggggctcgc gacggagttc     6180

gcccgcgagc ggggcgccgg gcacctctgg ctggaggtca ccaacgtcaa cgcaccggcg     6240

atccacgcgt accggcggat ggggttcacc ctctgcggcc tggacaccgc cctgtacgac     6300

ggcaccgcct cggacggcga gcaggcgctc tacatgagca tgccctgccc ctaatcagta     6360

ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg     6420

tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca     6480

tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa     6540

tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga     6600

gctcataact tcgtataatg tatgctatac gaacggtaga attcgatatc agatccacta     6660

gtggcctcat cttgatttat gtcttgtttg aggagatttc ggataaatat tcttcgaggg     6720

gaaaatgtcg taaaaataaa aacaataaag aaacaaaacg attaaaagaa aaactcaacc     6780

atcctcaaaa gtcctcctct tttttttcac gtgctgcgct gatgtaagca gcaggacatg     6840

gcaggaagaa aatcggagaa ttgtggccgc gacacctgac aaacaatcct caggcaaatg     6900

taaaattgta catgaaaggc tgcgtagtca atgtcttaga aggccggccc gcatgatcca     6960

gaagcagacc ataaaaaaaa taaaaattcg cggtctggca gcaggcataa gatgcatagc     7020

gttatcctaa atgacatcac gatgataaat cctccgccgc atgatgcttt tgatttgcct     7080

aagggcctgc catcgtgctg gctcaaacta ttgagggtca acataccttg aaaatccaag     7140

taaaaggatg gatatcgtta tactaaaagc aacacagaaa aggtccacgt cagttccaca     7200

caataacatt tacgtagtgt tcacgcgaag cagttacatc                           7240


<210>  3
<211>  1875
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of Brick 1 comprising the 2 plasmid

<400>  3
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtggtgct ttttttgttt tttatgtctt cgagtcatgt aattagttat      120

gtcgtggcat gctagctccg gattatcgat gataagctgt caaacatgag aattaattcc      180

acggactata gactatacct agtatactcc gtctactgta cgatacactt ccgctcaggt      240

ccttgtcctt taacgaggcc ttaccactct tttgttactc tattgatcca gctcagcaaa      300

ggcagtgtga tctaagattc tatcttcgcg atgtagtaaa actagctaga ccgagaaaga      360

gactagaaat gcaaaaggca cttctacaat ggctgccatc attattatcc gatgtgacgc      420

tgcagcttct caatgatatt cgaatacgct ttgaggagat acagcctaat atccgacaaa      480

ctgttttaca gatttacgat cgtacttgtt acccatcatt gaattttgaa catccgaacc      540

tgggagtttt ccctgaaaca gatagtatat ttgaacctgt ataataatat atagtctagc      600

gctttacgga agacaatgta tgtatttcgg ttcctggaga aactattgca tctattgcat      660

aggtaatctt gcacgtcgca tccccggttc attttctgcg tttccatctt gcacttcaat      720

agcatatctt tgttaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga      780

gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg      840

cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa      900

cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg      960

caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa     1020

tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg     1080

tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa     1140

gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg     1200

tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta     1260

tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat     1320

tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat     1380

aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa     1440

tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag     1500

atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat     1560

atatagcaaa gagatacttt tgagcaatgt ttgtggaaag cggctgttct ttgaaaagat     1620

aatgtatgat tatgctttca ctcatattta tacagaaact tgatgttttc tttcgagtat     1680

atacaaggtg attacatgta cgtttgaagt acaactctag attttgtagt gccctcttgg     1740

gctagcggta aaggtgcgca ttttttcaca ccctacaatg ttctgttcaa aagattttgg     1800

tcaaacgctg tagaagtgaa agttggtgcg catgtttcgg cgttcgaaac ttctccgcag     1860

tgaaagataa atgat                                                      1875


<210>  4
<211>  2125
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of Brick 2 comprising the expression 
       cassette of Dasher GFP, flanked upstream by the S. cerevisae TDH3
       promoter, and flanked downstream by the S. cerevisiae ADH1 
       terminator sequence

<400>  4
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtggtgct ttttttgttt tttatgtctt cgagtcatgt aattagttat      120

gtctggccgg atcgatgtac acaaccgact gcacccaaac gaacacaaat cttagcagtg      180

cttagtcaaa aaattagcct tttaattctg ctgtaacccg tacatgccca aaataggggg      240

cgggttacac agaatatata acatcgtagg tgtctgggtg aacagtttat tcctggcatc      300

cactaaatat aatggagccc gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc      360

accaaaatat tgttttcttc accaaccatc agttcatagg tccattctct tagcgcaact      420

acagagaaca ggggcacaaa caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac      480

ctgcctggag taaatgatga cacaaggcaa ttgacccacg catgtatcta tctcattttc      540

ttacaccttc tattaccttc tgctctctct gatttggaaa aagctgaaaa aaaaggttga      600

aaccagttcc ctgaaattat tcccctactt gactaataag tatataaaga cggtaggtat      660

tgattgtaat tctgtaaatc tatttcttaa acttcttaaa ttctactttt atagttagtc      720

ttttttttag ttttaaaaca ccaagaactt agtttcgaat aaacacacat aaacaaacaa      780

aatgaccgca ctaacagaag gagctaaact attcgaaaag gagattcctt acattacaga      840

attagagggt gatgtcgaag gaatgaaatt cattatcaag ggcgagggta ctggtgacgc      900

tactaccggt acgattaaag caaagtacat ctgtacaaca ggtgaccttc ctgttccgtg      960

ggctactctg gtgagcactt tgtcttatgg agttcaatgt tttgctaaat acccttcgca     1020

cattaaagac tttttcaaaa gtgcaatgcc tgagggctat actcaggaga gaacaatatc     1080

tttcgaagga gatggtgtgt ataagactag ggctatggtc acgtatgaaa gaggatccat     1140

ctacaataga gtaactttaa ctggtgaaaa cttcaaaaag gacggtcaca tccttagaaa     1200

gaatgttgcc tttcaatgcc caccatccat cttgtacatt ttgccagaca cagttaacaa     1260

tggtatcaga gttgagttta accaagctta tgacatagag ggtgtcaccg aaaagttggt     1320

tacaaaatgt tcacagatga atcgtcccct ggcaggatca gctgccgtcc atatcccacg     1380

ttaccatcat atcacttatc ataccaagct gtccaaagat cgtgatgaga gaagggatca     1440

catgtgtttg gttgaagtgg taaaggccgt ggatttggat acttaccaag gttgataaag     1500

cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat     1560

acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt     1620

tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct     1680

ctaccggcat gccgagcaaa tgcctgcaaa tcgctcccca tttcacccaa ttgtagatat     1740

gctaactcca gcaatgagtt gatgaatctc ggtgtgtatt ttatgtcctc agaggacaac     1800

ctcagaaagc ctgtatgcga agccacaatc ctttccaaca gaccatacta agtgaattct     1860

ttgaaaagat aatgtatgat tatgctttca ctcatattta tacagaaact tgatgttttc     1920

tttcgagtat atacaaggtg attacatgta cgtttgaagt acaactctag attttgtagt     1980

gccctcttgg gctagcggta aaggtgcgca ttttttcaca ccctacaatg ttctgttcaa     2040

aagattttgg tcaaacgctg tagaagtgaa agttggtgcg catgtttcgg cgttcgaaac     2100

ttctccgcag tgaaagataa atgat                                           2125


<210>  5
<211>  1846
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of Brick 3 comprising the dominant 
       resistance marker KanMX

<400>  5
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtggtgct ttttttgttt tttatgtctt cgagtcatgt aattagttat      120

gtctgagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg      180

cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc      240

gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg      300

gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg      360

ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag      420

gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat      480

acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga ctcacgtttc      540

gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga      600

taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga      660

gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag      720

actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc      780

tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga      840

agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt      900

gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca      960

ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa     1020

tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga     1080

ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt     1140

aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat     1200

cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata     1260

tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt     1320

ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt     1380

ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc     1440

ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat     1500

cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtg     1560

tcgaaaacga gctcgaattc tttgaaaaga taatgtatga ttatgctttc actcatattt     1620

atacagaaac ttgatgtttt ctttcgagta tatacaaggt gattacatgt acgtttgaag     1680

tacaactcta gattttgtag tgccctcttg ggctagcggt aaaggtgcgc attttttcac     1740

accctacaat gttctgttca aaagattttg gtcaaacgct gtagaagtga aagttggtgc     1800

gcatgtttcg gcgttcgaaa cttctccgca gtgaaagata aatgat                    1846


<210>  6
<211>  2047
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of Brick 4 comprising the kanR gene, a 
       bacterial selection marker, and pUC ORI

<400>  6
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtggtgct ttttttgttt tttatgtctt cgagtcatgt aattagttat      120

gtctgcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg      180

ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg      240

ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac      300

gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg      360

gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct      420

ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg      480

tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct      540

gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac      600

tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt      660

tcttgaagtg gtgggctaac tacggctaca ctagaagaac agtatttggt atctgcgctc      720

tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca      780

ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat      840

ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg      900

taactcacgt taagggattt tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg      960

ctctgctttt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga     1020

ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg     1080

cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca     1140

atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga     1200

gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca     1260

acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt     1320

cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca     1380

ggaatcgagt gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa     1440

tcaggatatt cttctaatac ctggaacgct gtttttccgg ggatcgcagt ggtgagtaac     1500

catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagtggcat aaattccgtc     1560

agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt     1620

ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat     1680

tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt     1740

aatcgcggcc tcgacgtttc ccgttgaata tggctgaatt ctttgaaaag ataatgtatg     1800

attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg     1860

tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg     1920

taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc     1980

tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat     2040

aaatgat                                                               2047


<210>  7
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer gR1-HXT1-f

<400>  7
tgaaagataa atgatgttgt agtcagcgcc tctctgtttt agagctagaa atagcaagtt       60


<210>  8
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer gR2-HXT2-f

<400>  8
tgaaagataa atgattgggt taaccttagg acgtcgtttt agagctagaa atagcaagtt       60


<210>  9
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer gR3-HXT3-f

<400>  9
tgaaagataa atgatggtgg ctagtatttg tttcggtttt agagctagaa atagcaagtt       60


<210>  10
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer gR4-GAL2-f

<400>  10
tgaaagataa atgattctaa ctctgcctgg acggcgtttt agagctagaa atagcaagtt       60


<210>  11
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer gR1-HXT1-r

<400>  11
ttctagctct aaaacagaga ggcgctgact acaacatcat ttatctttca ctgcggagaa       60


<210>  12
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer gR2-HXT2-r

<400>  12
ttctagctct aaaacgacgt cctaaggtta acccaatcat ttatctttca ctgcggagaa       60


<210>  13
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer gR3-HXT3-r

<400>  13
ttctagctct aaaaccgaaa caaatactag ccaccatcat ttatctttca ctgcggagaa       60


<210>  14
<211>  60
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer gR4-GAL2-r

<400>  14
ttctagctct aaaacgccgt ccaggcagag ttagaatcat ttatctttca ctgcggagaa       60


<210>  15
<211>  1945
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 1

<400>  15
tgaaagataa atgatgttgt agtcagcgcc tctctgtttt agagctagaa atagcaagtt       60

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg gtgctttttt      120

tgttttttat gtcttcgagt catgtaatta gttatgtcgt ggcatgctag ctccggatta      180

tcgatgataa gctgtcaaac atgagaatta attccacgga ctatagacta tacctagtat      240

actccgtcta ctgtacgata cacttccgct caggtccttg tcctttaacg aggccttacc      300

actcttttgt tactctattg atccagctca gcaaaggcag tgtgatctaa gattctatct      360

tcgcgatgta gtaaaactag ctagaccgag aaagagacta gaaatgcaaa aggcacttct      420

acaatggctg ccatcattat tatccgatgt gacgctgcag cttctcaatg atattcgaat      480

acgctttgag gagatacagc ctaatatccg acaaactgtt ttacagattt acgatcgtac      540

ttgttaccca tcattgaatt ttgaacatcc gaacctggga gttttccctg aaacagatag      600

tatatttgaa cctgtataat aatatatagt ctagcgcttt acggaagaca atgtatgtat      660

ttcggttcct ggagaaacta ttgcatctat tgcataggta atcttgcacg tcgcatcccc      720

ggttcatttt ctgcgtttcc atcttgcact tcaatagcat atctttgtta acgaagcatc      780

tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa      840

tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag      900

aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca      960

aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa     1020

caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct     1080

aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga     1140

taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc     1200

tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg     1260

ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc     1320

atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa     1380

cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg     1440

ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta     1500

gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg     1560

gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc     1620

aatgtttgtg gaaagcggct gttctttgaa aagataatgt atgattatgc tttcactcat     1680

atttatacag aaacttgatg ttttctttcg agtatataca aggtgattac atgtacgttt     1740

gaagtacaac tctagatttt gtagtgccct cttgggctag cggtaaaggt gcgcattttt     1800

tcacacccta caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa gtgaaagttg     1860

gtgcgcatgt ttcggcgttc gaaacttctc cgcagtgaaa gataaatgat tgggttaacc     1920

ttaggacgtc gttttagagc tagaa                                           1945


<210>  16
<211>  2195
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 2

<400>  16
tgaaagataa atgattgggt taaccttagg acgtcgtttt agagctagaa atagcaagtt       60

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg gtgctttttt      120

tgttttttat gtcttcgagt catgtaatta gttatgtctg gccggatcga tgtacacaac      180

cgactgcacc caaacgaaca caaatcttag cagtgcttag tcaaaaaatt agccttttaa      240

ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat atataacatc      300

gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg agcccgcttt      360

ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt tcttcaccaa      420

ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc acaaacaggc      480

aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat gatgacacaa      540

ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta ccttctgctc      600

tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa attattcccc      660

tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt aaatctattt      720

cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta aaacaccaag      780

aacttagttt cgaataaaca cacataaaca aacaaaatga ccgcactaac agaaggagct      840

aaactattcg aaaaggagat tccttacatt acagaattag agggtgatgt cgaaggaatg      900

aaattcatta tcaagggcga gggtactggt gacgctacta ccggtacgat taaagcaaag      960

tacatctgta caacaggtga ccttcctgtt ccgtgggcta ctctggtgag cactttgtct     1020

tatggagttc aatgttttgc taaataccct tcgcacatta aagacttttt caaaagtgca     1080

atgcctgagg gctatactca ggagagaaca atatctttcg aaggagatgg tgtgtataag     1140

actagggcta tggtcacgta tgaaagagga tccatctaca atagagtaac tttaactggt     1200

gaaaacttca aaaaggacgg tcacatcctt agaaagaatg ttgcctttca atgcccacca     1260

tccatcttgt acattttgcc agacacagtt aacaatggta tcagagttga gtttaaccaa     1320

gcttatgaca tagagggtgt caccgaaaag ttggttacaa aatgttcaca gatgaatcgt     1380

cccctggcag gatcagctgc cgtccatatc ccacgttacc atcatatcac ttatcatacc     1440

aagctgtcca aagatcgtga tgagagaagg gatcacatgt gtttggttga agtggtaaag     1500

gccgtggatt tggatactta ccaaggttga taaagcgaat ttcttatgat ttatgatttt     1560

tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt     1620

tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca     1680

ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgccga gcaaatgcct     1740

gcaaatcgct ccccatttca cccaattgta gatatgctaa ctccagcaat gagttgatga     1800

atctcggtgt gtattttatg tcctcagagg acaacctcag aaagcctgta tgcgaagcca     1860

caatcctttc caacagacca tactaagtga attctttgaa aagataatgt atgattatgc     1920

tttcactcat atttatacag aaacttgatg ttttctttcg agtatataca aggtgattac     1980

atgtacgttt gaagtacaac tctagatttt gtagtgccct cttgggctag cggtaaaggt     2040

gcgcattttt tcacacccta caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa     2100

gtgaaagttg gtgcgcatgt ttcggcgttc gaaacttctc cgcagtgaaa gataaatgat     2160

ggtggctagt atttgtttcg gttttagagc tagaa                                2195


<210>  17
<211>  1916
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 3

<400>  17
tgaaagataa atgatggtgg ctagtatttg tttcggtttt agagctagaa atagcaagtt       60

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg gtgctttttt      120

tgttttttat gtcttcgagt catgtaatta gttatgtctg agatctgttt agcttgcctc      180

gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct      240

tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc      300

ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta      360

cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg      420

tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt      480

ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca      540

taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg      600

gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca      660

atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt      720

agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg      780

cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact      840

gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat      900

attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt      960

ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt     1020

ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg     1080

aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc     1140

tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga     1200

gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt     1260

tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat     1320

aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga     1380

ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt     1440

aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga     1500

agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac     1560

tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa aacgagctcg aattctttga     1620

aaagataatg tatgattatg ctttcactca tatttataca gaaacttgat gttttctttc     1680

gagtatatac aaggtgatta catgtacgtt tgaagtacaa ctctagattt tgtagtgccc     1740

tcttgggcta gcggtaaagg tgcgcatttt ttcacaccct acaatgttct gttcaaaaga     1800

ttttggtcaa acgctgtaga agtgaaagtt ggtgcgcatg tttcggcgtt cgaaacttct     1860

ccgcagtgaa agataaatga ttctaactct gcctggacgg cgttttagag ctagaa         1916


<210>  18
<211>  2127
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 4

<400>  18
cgcagtgaaa gataaatgat tctaactctg cctggacggc gttttagagc tagaaatagc       60

aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct      120

ttttttgttt tttatgtctt cgagtcatgt aattagttat gtctgcgagc ggtatcagct      180

cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg      240

tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc      300

cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga      360

aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct      420

cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg      480

gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag      540

ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat      600

cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac      660

aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtgggctaac      720

tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc      780

ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt      840

tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc      900

ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt      960

tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgctttt agaaaaactc     1020

atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg     1080

aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag     1140

atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc     1200

ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga     1260

gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc     1320

gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag     1380

gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgagt gcaaccggcg     1440

caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac     1500

ctggaacgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg     1560

gataaaatgc ttgatggtcg gaagtggcat aaattccgtc agccagttta gtctgaccat     1620

ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc     1680

atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc     1740

ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc     1800

ccgttgaata tggctgaatt ctttgaaaag ataatgtatg attatgcttt cactcatatt     1860

tatacagaaa cttgatgttt tctttcgagt atatacaagg tgattacatg tacgtttgaa     1920

gtacaactct agattttgta gtgccctctt gggctagcgg taaaggtgcg cattttttca     1980

caccctacaa tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg aaagttggtg     2040

cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat aaatgatgtt gtagtcagcg     2100

cctctctgtt ttagagctag aaatagc                                         2127


<210>  19
<211>  382
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor DNA used in the CRISPR-CAS9-mediated deletion of genes 
       HXT5-HXT1-HXT4 in S. cerevisiae strain BIE272

<400>  19
tagtcgaacg gttctccctc taagaataag aaactatcaa aagaaagaga aaagtcgatt       60

gaataatttt tctatatata atatacgcaa acaagattcg ctttcacttt gcaattttac      120

ttcatagctt tgttaaaacc agcaaaaaat attatttttc tagaaaaaag aatatattag      180

aggtaaagaa agatcagtta attaaataat atagacttcc ttcaaggaat aaaaataata      240

tttgatcttc aatttttaac gctttatgaa atatatattt atgatttaat agtttataga      300

taatacttta tcaaactaca attttgaatt atgttttttg tttcatagtt ctggaagagg      360

ttaggaatag gccgaattct cc                                               382


<210>  20
<211>  468
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor DNA used in the CRISPR-CAS9-mediated deletion of gene HXT2 
       in S. cerevisiae strain BIE272

<400>  20
cggtacgccg cctaacgata gtaataggcc acgcaactgg cgtggacgac aacaataagt       60

cgcccatttt ttatgttttc aaaacctagc aacccccacc aaacttgtca tcgttcccgg      120

attcacaaat gatataaaaa gcgattacaa ttctacattc taaccagatt tgagatttcc      180

tctttctcaa ttcctcttat attagattat aagaacaaca aattaaatta caaaaagact      240

tataaagcaa catagagatt atacttaaac tagcactgat ttttttaagg ctaatggcta      300

ctaatacttt aatagatgat cttcatactt ttttatttaa cgatttttaa tgatgttttt      360

atttgtacca ctcatttatc tagatttttt taatactgat caaatcttac ggactcgacg      420

ttaaaaagtt cctacatacg tctggtactt gaaacgctgc ttcgaggt                   468


<210>  21
<211>  422
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor DNA used in the CRISPR-CAS9-mediated deletion of genes 
       HXT3-HXT6-HXT7 in S. cerevisiae strain BIE272

<400>  21
gactatatcc ccagagaatt tctccagaag agaagaaaag tcaaagtttt ttttcgcttg       60

ggggttgcat ataaatacag gcgctgtttt atcttcagca tgaatattcc ataattttac      120

ttaatagctt ttcataaata atagaatcac aaacaaaatt tacatctgag ttaaacaatc      180

tttgcgaaca cttttattaa ttcatgatca cgctctaatt tgtgcatttg aaatgtactc      240

taattctaat tttatatttt taatgatatc ttgaaaagta aatacgtttt taatatatac      300

aaaataatac agtttaattt tcaagttttt gatcatttgt tctcagaaag ttgagtggga      360

cggagacaaa gaaactttaa agagaaatgc aaagtgggaa gaagtcagtt gtttaccgac      420

cg                                                                     422


<210>  22
<211>  414
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor DNA used in the CRISPR-CAS9-mediated deletion of gene GAL2 
       in S. cerevisiae strain BIE272

<400>  22
tggagatagg attagttttg tagacatata taaacaatca gtaattggat tgaaaatttg       60

gtgttgtgaa ttgctcttca ttatgcacct tattcaatta tcatcaagaa tagcaatagt      120

taagtaaaca caagattaac ataataaaaa aaataattct ttcataatgg cagttgagga      180

gaacaatatg cctgttgttt cacagcaacc ccaagctggt gaagacgtga cctatcgtta      240

gaagaaattc aagaattatg ggaagaaggt gttttacctt ggaaatctga aggctggatt      300

ccttcatcca gaagaggtaa taattacgat ttagaggatt tacaacatga cgacaaaccg      360

tggtacaagg ccatgctaga ataatgcgtt tgaagtgaga cgctccatca tctc            414


<210>  23
<211>  11742
<212>  DNA
<213>  artificial sequence

<220>
<223>  CAS9 expression vector pCSN061

<400>  23
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca       60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg      120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc      180

accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat      240

atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa      300

aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa      360

gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat      420

tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta      480

atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg      540

cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa      600

agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa      660

atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg      720

ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt      780

ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag      840

actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg      900

attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg      960

gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa     1020

atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg     1080

agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat     1140

aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga     1200

aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt     1260

ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa     1320

atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca     1380

gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc     1440

gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg     1500

aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg     1560

ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg     1620

gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg     1680

ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga     1740

tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga     1800

ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag     1860

cgcgcgtaat acgactcact atagggcgaa ttgggtacct tttctttttt tgcggtcacc     1920

cccatgtggc ggggaggcag aggagtaggt agagcaacga atcctactat ttatccaaat     1980

tagtctagga actctttttc tagatttttt agatttgagg gcaagcgctg ttaacgactc     2040

agaaatgtaa gcactacgga gtagaacgag aaatccgcca taggtggaaa tcctagcaaa     2100

atcttgctta ccctagctag cctcaggtaa gctagcctta gcctgtcaaa tttttttcaa     2160

aatttggtaa gtttctacta gcaaagcaaa cacggttcaa caaaccgaaa actccactca     2220

ttatacgtgg aaaccgaaac aaaaaaacaa aaaccaaaat actcgccaat gagaaagttg     2280

ctgcgtttct actttcgagg aagaggaact gagaggattg actacgaaag gggcaaaaac     2340

gagtcgtatt ctcccattat tgtctgctac cacgcggtct agtagaataa gcaaccagtc     2400

aacgctaaga caggtaatca aaataccagt ctgctggcta cgggctagtt tttacctctt     2460

ttagaaccca ctgtaaaagt ccgttgtaaa gcccgttctc actgttggcg tttttttttt     2520

tttggtttag tttcttattt ttcatttttt tctttcatga ccaaaaacaa acaaatctcg     2580

cgatttgtac tgcggccact ggggcgtggc caaaaaaatg acaaatttag aaaccttagt     2640

ttctgatttt tcctgttatg aggagatatg ataaaaaata ttactgcttt attgtttttt     2700

ttttatctac tgaaatagag aaacttaccc aaggaggagg caaaaaaaag agtatatata     2760

cagcagctac cattcagatt ttaatatatt cttttctctt cttctacact attattataa     2820

taattttact atattcattt ttagcttaaa acctcataga atattattct tcagtcactc     2880

gcttaaatac ttatcaaaaa tggacaagaa atactctatt ggtttggata tcgggaccaa     2940

ctccgtcggt tgggctgtca tcaccgacga atacaaggtt ccatccaaga aattcaaggt     3000

cttgggtaac actgacagac actctatcaa gaagaatttg atcggtgctt tgttgttcga     3060

ctccggtgaa accgctgaag ctaccagatt gaagcgtacc gctcgtcgta gatacactag     3120

acgtaaaaac cgtatttgtt acttgcaaga aatcttttct aacgaaatgg ccaaggttga     3180

cgactctttc ttccacagat tggaagaatc tttcttggtt gaagaagaca agaagcacga     3240

aagacatcca atcttcggta acatcgttga cgaagttgct taccacgaaa aataccctac     3300

catctaccat ttgagaaaga agttggtcga ttccaccgac aaggctgatt tgagattgat     3360

ctatttggcc ttggctcaca tgatcaagtt cagaggtcac ttcttgattg aaggtgactt     3420

gaacccagac aactctgacg tcgacaaatt gttcatccaa ttggtccaaa cctacaacca     3480

attattcgag gaaaacccaa ttaacgcttc tggtgttgat gctaaggcca tcttatctgc     3540

ccgtttgtcc aagtctagac gtttggaaaa cttgattgct caattgcctg gtgaaaagaa     3600

aaacggtttg ttcggtaact tgatcgcttt gtccttgggt ttgaccccaa acttcaagtc     3660

caacttcgac ttggctgaag atgccaagtt gcaattgtcc aaggacacct acgacgacga     3720

cttagacaac ttgttggctc aaatcggtga ccaatacgcc gacttgttct tggctgccaa     3780

aaacttatct gacgctatct tgttgtctga catcttgaga gttaacactg aaattaccaa     3840

ggctccattg tctgcttcta tgatcaaaag atacgacgaa caccaccaag atctgacttt     3900

gttgaaggct ttggttagac aacaattgcc agaaaagtac aaggaaatct tcttcgacca     3960

atccaaaaat ggttacgccg gttacattga cggtggtgct tctcaggaag aattctacaa     4020

gttcatcaag ccaattttgg aaaagatgga tggtactgaa gaattattgg ttaagttgaa     4080

cagagaagac ttattgagaa agcaacgtac cttcgataac ggttctatcc cacaccaaat     4140

ccacttgggt gaattgcacg ccattttgag aagacaggaa gatttctatc cattcctaaa     4200

ggacaacaga gaaaagatcg aaaagatctt aactttcaga atcccatact acgtcggtcc     4260

attggccaga ggtaattcta gattcgcttg gatgaccaga aagtctgaag aaaccatcac     4320

cccatggaac ttcgaagaag tcgtcgacaa gggtgcttct gcccaatctt tcatcgaaag     4380

aatgaccaac tttgataaga acttgccaaa cgagaaggtc ttgccaaagc actctttgtt     4440

gtacgaatac ttcaccgtct acaacgaatt aaccaaggtt aaatacgtta ctgaaggtat     4500

gagaaagcca gctttcctat ccggtgaaca aaagaaggct attgttgact tgttgtttaa     4560

gaccaacaga aaggtcactg ttaagcaatt gaaggaagac tacttcaaga agattgaatg     4620

tttcgattcc gtcgaaatct ccggtgttga agaccgtttc aatgcttctt tgggcaccta     4680

ccacgatttg ttaaagatca tcaaggacaa ggacttttta gataacgaag aaaacgaaga     4740

catcttggaa gatatcgttt tgaccttgac tcttttcgag gacagagaaa tgattgaaga     4800

gagattgaag acctacgctc acttgttcga cgataaagtt atgaagcaac taaagagaag     4860

aagatacact ggttggggta gattgtccag aaagttgatt aacggtatca gagacaagca     4920

atccggtaag actattttag actttttgaa atccgatggt ttcgctaaca gaaactttat     4980

gcaattgatt cacgacgatt ctttgacttt caaggaagac attcaaaaag cccaagtctc     5040

tggtcaaggt gattctttgc acgaacacat cgctaacttg gctggttctc cagctattaa     5100

gaagggtatc ttacaaaccg tcaaggtcgt tgatgaattg gtcaaagtca tgggtagaca     5160

caagccagaa aatattgtca tcgaaatggc tagagaaaac caaactactc aaaagggtca     5220

aaagaactct agagaacgta tgaagagaat tgaagaaggt atcaaggagt tgggttctca     5280

aattttgaaa gaacacccag tcgaaaacac tcaattacaa aacgaaaagc tatacttgta     5340

ctacttgcaa aacggtcgtg acatgtacgt cgaccaagaa ttggatatca acagattgtc     5400

tgactacgat gtcgatcata tcgtcccaca atcgttcttg aaggacgatt ccattgacaa     5460

caaagttttg actagatctg acaagaacag aggtaagtct gataacgttc catctgaaga     5520

agttgttaag aagatgaaga actactggag acaattgttg aatgctaagt tgatcactca     5580

aagaaagttc gacaacttga ccaaggctga aagaggtggt ttgtccgaat tggacaaagc     5640

cggtttcatc aagagacaat tagtcgaaac tagacaaatc accaagcatg ttgctcaaat     5700

cttggattcc agaatgaaca ctaagtacga tgaaaacgac aaactaatta gagaagttaa     5760

ggtcatcact ttgaagtcta agttggtttc tgacttcaga aaggacttcc aattttacaa     5820

ggtcagagaa atcaacaact accatcacgc tcacgatgcc tacttgaacg ctgttgtcgg     5880

tactgcctta atcaaaaagt acccaaagtt ggaatctgaa ttcgtttacg gtgactacaa     5940

ggtttacgat gttagaaaga tgatcgccaa gtctgaacaa gaaattggta aggccactgc     6000

taagtacttc ttctactcta acatcatgaa ctttttcaag actgaaatca ctttagctaa     6060

cggtgaaatt agaaagcgtc cattgattga aaccaatggt gaaactggtg aaattgtctg     6120

ggacaagggt agagatttcg ctaccgtcag aaaggttttg tctatgccac aagttaacat     6180

cgtcaagaag actgaagttc aaactggtgg tttctctaag gaatccattt tgccaaagag     6240

aaactctgac aagttgattg ctagaaagaa ggactgggat cctaagaagt acggtggttt     6300

cgactctcca actgttgctt actccgtttt ggtcgttgct aaggttgaaa agggtaagtc     6360

taagaagttg aagtctgtta aggaattgtt gggtatcacc atcatggaaa gatcctcctt     6420

cgaaaagaac ccaatcgact ttttggaagc taagggttac aaggaagtca agaaggattt     6480

gatcattaag ttaccaaaat actccttgtt cgaattggaa aacggtagaa agagaatgtt     6540

ggcctccgct ggtgaactac aaaaaggtaa cgaattggct ttaccatcta agtacgttaa     6600

cttcttgtac ttggcttccc actacgaaaa gttgaaaggt tccccagaag acaacgaaca     6660

aaagcaattg tttgttgaac aacacaagca ctacttggat gaaattattg aacaaatctc     6720

cgaattctcc aagagagtca ttttggctga tgctaactta gataaggttt tatccgctta     6780

caacaagcac agagacaaac caatcagaga acaagctgaa aacatcattc atttgttcac     6840

tttaaccaac ttgggtgctc cagctgcttt caaatacttc gacactacca ttgacagaaa     6900

gagatacact tccaccaaag aagttttaga tgctactttg attcaccaat ctattaccgg     6960

tttgtacgaa accagaattg acttgtctca attgggtggt gattccagag ctgatccaaa     7020

gaagaagaga aaggtgtaaa ggagttaaag gcaaagtttt cttttctaga gccgttccca     7080

caaataatta tacgtatatg cttcttttcg tttactatat atctatattt acaagccttt     7140

attcactgat gcaatttgtt tccaaatact tttttggaga tctcataact agatatcatg     7200

atggcgcaac ttggcgctat cttaattact ctggctgcca ggcccgtgta gagggccgca     7260

agaccttctg tacgccatat agtctctaag aacttgaaca agtttctaga cctattgccg     7320

cctttcggat cgctattgtt gcggccgcca gctgaagctt cgtacgctgc aggtcgacga     7380

attctaccgt tcgtataatg tatgctatac gaagttatag atctgtttag cttgcctcgt     7440

ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg     7500

acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc     7560

atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg     7620

gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc     7680

cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct     7740

ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata     7800

aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga     7860

tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat     7920

ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag     7980

cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc     8040

tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc     8100

gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat     8160

tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc     8220

ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt     8280

ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa     8340

agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc     8400

acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt     8460

cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc     8520

tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa     8580

attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt     8640

cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa     8700

tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag     8760

ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg     8820

ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctcata acttcgtata     8880

atgtatgcta tacgaacggt agaattcgaa tcagatccac tagtggccta tgcggccgcc     8940

accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta     9000

atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat     9060

aggagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt     9120

aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta     9180

atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc     9240

gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa     9300

ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa     9360

aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct     9420

ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac     9480

aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc     9540

gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc     9600

tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg     9660

tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga     9720

gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag     9780

cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta     9840

cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag     9900

agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg     9960

caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac    10020

ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc    10080

aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag    10140

tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc    10200

agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac    10260

gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc    10320

accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg    10380

tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag    10440

tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc    10500

acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac    10560

atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag    10620

aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac    10680

tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg    10740

agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc    10800

gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact    10860

ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg    10920

atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa    10980

tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt    11040

tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg    11100

tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg    11160

gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat    11220

aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag    11280

atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct    11340

ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc    11400

ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct    11460

tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta    11520

tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat    11580

acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga    11640

tacgaggcgc gtgtaagtta caggcaagcg atccgtccta agaaaccatt attatcatga    11700

cattaaccta taaaaatagg cgtatcacga ggccctttcg tc                       11742


<210>  24
<211>  1609
<212>  DNA
<213>  artificial sequence

<220>
<223>  Brick 3b (natMX-fragment)

<400>  24
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtggtgct ttttttgttt tttatgtctt cgagtcatgt aattagttat      120

gtctgagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg      180

cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc      240

gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg      300

gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg      360

ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag      420

gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat      480

acagttctca catcacatcc gaacataaac aaccatgggt accactcttg acgacacggc      540

ttaccggtac cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc      600

cttcaccacc gacaccgtct tccgcgtcac cgccaccggg gacggcttca ccctgcggga      660

ggtgccggtg gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc      720

ggacgacggg gaggacggcg acccggactc ccggacgttc gtcgcgtacg gggacgacgg      780

cgacctggcg ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga      840

ggacatcgag gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct      900

cgcgacggag ttcgcccgcg agcggggcgc cgggcacctc tggctggagg tcaccaacgt      960

caacgcaccg gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac     1020

cgccctgtac gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg     1080

cccctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag     1140

tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt     1200

cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc     1260

aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca     1320

gtgtcgaaaa cgagctcgaa ttctttgaaa agataatgta tgattatgct ttcactcata     1380

tttatacaga aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg     1440

aagtacaact ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt     1500

cacaccctac aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg     1560

tgcgcatgtt tcggcgttcg aaacttctcc gcagtgaaag ataaatgat                 1609


<210>  25
<211>  61
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer g1-INT59-f

<400>  25
tgaaagataa atgatcagaa aactcttagc ttttccgttt tagagctaga aatagcaagt       60

t                                                                       61


<210>  26
<211>  61
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer g2-YPRc-f

<400>  26
tgaaagataa atgatccaat atggtatgcc gagtctgttt tagagctaga aatagcaagt       60

t                                                                       61


<210>  27
<211>  61
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer g3-INT1-f

<400>  27
tgaaagataa atgatctatt agaaccaggg aggtccgttt tagagctaga aatagcaagt       60

t                                                                       61


<210>  28
<211>  50
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer Brick4_to3-f

<400>  28
gccatccagt gtcgaaaacg agctcgcgag cggtatcagc tcactcaaag                  50


<210>  29
<211>  61
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer g1-INT59-r

<400>  29
ttctagctct aaaacggaaa agctaagagt tttctgatca tttatctttc actgcggaga       60

a                                                                       61


<210>  30
<211>  61
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer g2-YPRc-r

<400>  30
ttctagctct aaaacagact cggcatacca tattggatca tttatctttc actgcggaga       60

a                                                                       61


<210>  31
<211>  61
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer g3-INT1-r

<400>  31
ttctagctct aaaacggacc tccctggttc taatagatca tttatctttc actgcggaga       60

a                                                                       61


<210>  32
<211>  50
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer Brick 3_to4-r

<400>  32
ctttgagtga gctgataccg ctcgcgagct cgttttcgac actggatggc                  50


<210>  33
<211>  1945
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 1 (Example 4)

<400>  33
tgaaagataa atgatagaaa actcttagct tttccgtttt agagctagaa atagcaagtt       60

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg gtgctttttt      120

tgttttttat gtcttcgagt catgtaatta gttatgtcgt ggcatgctag ctccggatta      180

tcgatgataa gctgtcaaac atgagaatta attccacgga ctatagacta tacctagtat      240

actccgtcta ctgtacgata cacttccgct caggtccttg tcctttaacg aggccttacc      300

actcttttgt tactctattg atccagctca gcaaaggcag tgtgatctaa gattctatct      360

tcgcgatgta gtaaaactag ctagaccgag aaagagacta gaaatgcaaa aggcacttct      420

acaatggctg ccatcattat tatccgatgt gacgctgcag cttctcaatg atattcgaat      480

acgctttgag gagatacagc ctaatatccg acaaactgtt ttacagattt acgatcgtac      540

ttgttaccca tcattgaatt ttgaacatcc gaacctggga gttttccctg aaacagatag      600

tatatttgaa cctgtataat aatatatagt ctagcgcttt acggaagaca atgtatgtat      660

ttcggttcct ggagaaacta ttgcatctat tgcataggta atcttgcacg tcgcatcccc      720

ggttcatttt ctgcgtttcc atcttgcact tcaatagcat atctttgtta acgaagcatc      780

tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa      840

tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag      900

aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca      960

aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa     1020

caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct     1080

aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga     1140

taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc     1200

tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg     1260

ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc     1320

atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa     1380

cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg     1440

ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta     1500

gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg     1560

gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc     1620

aatgtttgtg gaaagcggct gttctttgaa aagataatgt atgattatgc tttcactcat     1680

atttatacag aaacttgatg ttttctttcg agtatataca aggtgattac atgtacgttt     1740

gaagtacaac tctagatttt gtagtgccct cttgggctag cggtaaaggt gcgcattttt     1800

tcacacccta caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa gtgaaagttg     1860

gtgcgcatgt ttcggcgttc gaaacttctc cgcagtgaaa gataaatgat caatatggta     1920

tgccgagtct gttttagagc tagaa                                           1945


<210>  34
<211>  2195
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 2 (Example 4)

<400>  34
tgaaagataa atgatcaata tggtatgccg agtctgtttt agagctagaa atagcaagtt       60

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg gtgctttttt      120

tgttttttat gtcttcgagt catgtaatta gttatgtctg gccggatcga tgtacacaac      180

cgactgcacc caaacgaaca caaatcttag cagtgcttag tcaaaaaatt agccttttaa      240

ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat atataacatc      300

gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg agcccgcttt      360

ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt tcttcaccaa      420

ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc acaaacaggc      480

aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat gatgacacaa      540

ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta ccttctgctc      600

tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa attattcccc      660

tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt aaatctattt      720

cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta aaacaccaag      780

aacttagttt cgaataaaca cacataaaca aacaaaatga ccgcactaac agaaggagct      840

aaactattcg aaaaggagat tccttacatt acagaattag agggtgatgt cgaaggaatg      900

aaattcatta tcaagggcga gggtactggt gacgctacta ccggtacgat taaagcaaag      960

tacatctgta caacaggtga ccttcctgtt ccgtgggcta ctctggtgag cactttgtct     1020

tatggagttc aatgttttgc taaataccct tcgcacatta aagacttttt caaaagtgca     1080

atgcctgagg gctatactca ggagagaaca atatctttcg aaggagatgg tgtgtataag     1140

actagggcta tggtcacgta tgaaagagga tccatctaca atagagtaac tttaactggt     1200

gaaaacttca aaaaggacgg tcacatcctt agaaagaatg ttgcctttca atgcccacca     1260

tccatcttgt acattttgcc agacacagtt aacaatggta tcagagttga gtttaaccaa     1320

gcttatgaca tagagggtgt caccgaaaag ttggttacaa aatgttcaca gatgaatcgt     1380

cccctggcag gatcagctgc cgtccatatc ccacgttacc atcatatcac ttatcatacc     1440

aagctgtcca aagatcgtga tgagagaagg gatcacatgt gtttggttga agtggtaaag     1500

gccgtggatt tggatactta ccaaggttga taaagcgaat ttcttatgat ttatgatttt     1560

tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt     1620

tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca     1680

ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgccga gcaaatgcct     1740

gcaaatcgct ccccatttca cccaattgta gatatgctaa ctccagcaat gagttgatga     1800

atctcggtgt gtattttatg tcctcagagg acaacctcag aaagcctgta tgcgaagcca     1860

caatcctttc caacagacca tactaagtga attctttgaa aagataatgt atgattatgc     1920

tttcactcat atttatacag aaacttgatg ttttctttcg agtatataca aggtgattac     1980

atgtacgttt gaagtacaac tctagatttt gtagtgccct cttgggctag cggtaaaggt     2040

gcgcattttt tcacacccta caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa     2100

gtgaaagttg gtgcgcatgt ttcggcgttc gaaacttctc cgcagtgaaa gataaatgat     2160

tattagaacc agggaggtcc gttttagagc tagaa                                2195


<210>  35
<211>  1397
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 3b (Example 4)

<400>  35
tgaaagataa atgattatta gaaccaggga ggtccgtttt agagctagaa atagcaagtt       60

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg gtgctttttt      120

tgttttttat gtcttcgagt catgtaatta gttatgtctg agatctgttt agcttgcctc      180

gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct      240

tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc      300

ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta      360

cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg      420

tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt      480

ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca      540

taaacaacca tgggtaccac tcttgacgac acggcttacc ggtaccgcac cagtgtcccg      600

ggggacgccg aggccatcga ggcactggat gggtccttca ccaccgacac cgtcttccgc      660

gtcaccgcca ccggggacgg cttcaccctg cgggaggtgc cggtggaccc gcccctgacc      720

aaggtgttcc ccgacgacga atcggacgac gaatcggacg acggggagga cggcgacccg      780

gactcccgga cgttcgtcgc gtacggggac gacggcgacc tggcgggctt cgtggtcgtc      840

tcgtactccg gctggaaccg ccggctgacc gtcgaggaca tcgaggtcgc cccggagcac      900

cgggggcacg gggtcgggcg cgcgttgatg gggctcgcga cggagttcgc ccgcgagcgg      960

ggcgccgggc acctctggct ggaggtcacc aacgtcaacg caccggcgat ccacgcgtac     1020

cggcggatgg ggttcaccct ctgcggcctg gacaccgccc tgtacgacgg caccgcctcg     1080

gacggcgagc aggcgctcta catgagcatg ccctgcccct aatcagtact gacaataaaa     1140

agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat     1200

tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg     1260

cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta     1320

tactgctgtc gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgcgagcgg     1380

tatcagctca ctcaaag                                                    1397


<210>  36
<211>  1983
<212>  DNA
<213>  artificial sequence

<220>
<223>  extended Brick 4 (Example 4)

<400>  36
gccatccagt gtcgaaaacg agctcgcgag cggtatcagc tcactcaaag gcggtaatac       60

ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa      120

aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg      180

acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa      240

gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc      300

ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac      360

gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac      420

cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg      480

taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt      540

atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa      600

cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct      660

cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga      720

ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg      780

ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcttgcgccg      840

tcccgtcaag tcagcgtaat gctctgcttt tagaaaaact catcgagcat caaatgaaac      900

tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat      960

gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg     1020

attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta     1080

tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc     1140

atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca     1200

tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg     1260

ttaaaaggac aattacaaac aggaatcgag tgcaaccggc gcaggaacac tgccagcgca     1320

tcaacaatat tttcacctga atcaggatat tcttctaata cctggaacgc tgtttttccg     1380

gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc     1440

ggaagtggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg     1500

gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag     1560

cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa     1620

tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctgaat     1680

tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt     1740

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt     1800

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt     1860

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga     1920

aacttctccg cagtgaaaga taaatgatag aaaactctta gcttttccgt tttagagcta     1980

gaa                                                                   1983


<210>  37

<400>  37
000

<210>  38
<211>  2540
<212>  DNA
<213>  artificial sequence

<220>
<223>  con5 - Low p (KlTDH2p) - crtE - ScTDH3t - conA expression 
       cassette

<400>  38
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt gtgccgtaaa       60

aactaaaacg agcccccacc aaagaacaaa aaagaaggtg ctgggccccc actttcttcc      120

cttgcacgtg ataggaagat ggctacagaa acaagaagat ggaaatcgaa ggaaagaggg      180

agactggaag ctgtaaaaac tgaaatgaaa aaaaaaaaaa aaaaaaaaaa caagaagctg      240

aaaatggaag actgaaattt gaaaaatggt aaaaaaaaaa aagaaacacg aagctaaaaa      300

cctggattcc attttgagaa gaagcaagaa aggtaagtat ggtaacgacc gtacaggcaa      360

gcgcgaaggc aaatggaaaa gctggagtcc ggaagataat catttcatct tcttttgtta      420

gaacagaaca gtggatgtcc ctcatctcgg taacgtattg tccatgccct agaactctct      480

gtccctaaaa agaggacaaa aacccaatgg tttccccagc ttccagtgga gccaccgatc      540

ccactggaaa ccactggaca ggaagagaaa atcacggact tcctctattg aaggataatt      600

caacactttc accagatccc aaatgtcccg cccctattcc cgtgttccat cacgtaccat      660

aacttaccat ttcatcacgt tctctatggc acactggtac tgcttcgact gctttgcttc      720

atcttctcta tgggccaatg agctaatgag cacaatgtgc tgcgaaataa agggatatct      780

aatttatatt attacattat aatatgtact agtgtggtta ttggtaattg tacttaattt      840

tgatatataa agggtggatc tttttcattt tgaatcagaa ttggaattgc aacttgtctc      900

ttgtcactat tacttaatag taattatatt tcttattaac ctttttttta agtcaaaaca      960

ccaaggacaa gaactactct tcaaaggtat ttcaagttat catacgtgtc acacacgctt     1020

cacagtttca agtaaaaaaa aagaatatta cacaatggac tacgctaaca tcttgactgc     1080

cattcctttg gaattcaccc cacaagatga cattgtcttg ttggaaccat accactactt     1140

aggtaagaac ccaggtaagg aaatcagatc tcaattgatt gaagctttca actactggtt     1200

agatgtcaag aaggaagact tggaagttat ccaaaatgtt gttggtatgt tgcacaccgc     1260

ttctttgttg atggatgatg ttgaagattc ttccgtcttg agaagaggtt ctccagttgc     1320

tcatttgatc tacggtattc cacaaaccat caacactgct aactacgttt acttcttggc     1380

ttaccaagaa atcttcaaat tgcgtccaac tccaattcca atgccagtta tcccaccatc     1440

ttctgcttct ttgcaatctt ctgtctcctc cgcctcctct tcctcttctg cctcctctga     1500

aaacggtggt acctccactc caaactccca aatcccattc tccaaggaca cctacttgga     1560

caaggttatc actgacgaaa tgttgtcttt gcaccgtggt caaggtttgg aattattctg     1620

gagagactct ttgacctgtc catctgaaga agaatacgtc aagatggtct tgggtaagac     1680

cggtggtttg ttcagaattg ctgtcagatt gatgatggcc aagtctgaat gtgacattga     1740

ctttgttcaa ttggttaact tgatttccat ctacttccaa atcagagatg actacatgaa     1800

cttgcaatcc tctgaatacg ctcacaacaa gaacttcgct gaagacttga ctgaaggtaa     1860

gttctccttc ccaaccattc actccattca cgctaaccca tcttccagat tggttatcaa     1920

cactttacaa aagaagtcca cttctccaga aatcttacat cactgtgtca actacatgag     1980

aactgaaacc cactctttcg aatacactca agaagtcttg aacactttat ctggtgcttt     2040

ggaaagagaa ttgggtagat tacaaggtga atttgctgaa gctaactcca agatcgattt     2100

gggtgacgtt gaatctgaag gtagaaccgg taagaacgtc aaattggaag ccatcttgaa     2160

gaaattggct gatatccctc tataaagtga atttacttta aatcttgcat ttaaataaat     2220

tttcttttta tagctttatg acttagtttc aatttatata ctattttaat gacattttcg     2280

attcattgat tgaaagcttt gtgttttttc ttgatgcgct attgcattgt tcttgtcttt     2340

ttcgccacat gtaatatctg tagtagatac ctgatacatt gtggatgctg agtgaaattt     2400

tagttaataa tggaggcgct cttaataatt ttggggatat tggctttttt ttttaaagtt     2460

tacaaatgaa ttttttccgc caggatcctc ttgcccatcg aacgtacaag tactcctctg     2520

ttctctcctt cctttgcttt                                                 2540


<210>  39
<211>  2540
<212>  DNA
<213>  artificial sequence

<220>
<223>  con5 - Med p (KlPGK1p) - crtE - ScTDH3t - conA expression 
       cassette

<400>  39
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt gtgcgttcct       60

catcactaga agccgaactg ttgtcttcag tggggattgg ttcgacattt tgccaattgc      120

tgtcgatgta ccctttcaaa gccatgtacc ttaaatcttc atccttggca agtagattca      180

tcgggtgtgt ttgaagtaag aatatttgct tgtttttatg gtatcaaagg tatatgttgt      240

agaagacaat ttccggtaat ccaattgtct gtctgctcag tttagcacat gtatagtacg      300

ttgcacatag tctacaatat tcagcattca gcattcagta tacagcatat ggctaaatga      360

tcacaaatgt gattgatgat ttgacacgac tagaaaagag aacgaaaaag ggaaattcca      420

tgtcacgtgc gttggcacgt gacatggaat atcgaagaaa gaaaaaaaaa acgatctcgt      480

cctagtggaa gcccagagtc tggtcccccc ggagtcttcc caaaacaaga agctgacaca      540

tgttgacaca gaacacccca cagcaaatgc accacgctac gtagatcagg aagcttaact      600

ctagcgacct gtcgctcgcc ccacagaacc tcacccgaga accacacatt acacgccgcc      660

agctcccact atactcatct tgcttccctt aagcgttctc acgattcgtt cgctgccctt      720

cttcaagagt cttctgattc taattctcat tcgaaatcct ctacagttaa tgaattgctt      780

gacatgacat tcattgtctc atggttttgg ctttttggct tttgtctttt aaagctatat      840

caactttaca tataaatata cgtcaaaagg ggattcatta attagaaaat tctctttttc      900

aatagttgct attcattatc aatctattca actcaattgg ttattatttt catctttttg      960

tcatcctaaa ccatcaacaa tatttaaata tatctgttgc tacattaaga gttacttcag     1020

aaataacaaa aaaatcgatc aagaattaat aaaaatggac tacgctaaca tcttgactgc     1080

cattcctttg gaattcaccc cacaagatga cattgtcttg ttggaaccat accactactt     1140

aggtaagaac ccaggtaagg aaatcagatc tcaattgatt gaagctttca actactggtt     1200

agatgtcaag aaggaagact tggaagttat ccaaaatgtt gttggtatgt tgcacaccgc     1260

ttctttgttg atggatgatg ttgaagattc ttccgtcttg agaagaggtt ctccagttgc     1320

tcatttgatc tacggtattc cacaaaccat caacactgct aactacgttt acttcttggc     1380

ttaccaagaa atcttcaaat tgcgtccaac tccaattcca atgccagtta tcccaccatc     1440

ttctgcttct ttgcaatctt ctgtctcctc cgcctcctct tcctcttctg cctcctctga     1500

aaacggtggt acctccactc caaactccca aatcccattc tccaaggaca cctacttgga     1560

caaggttatc actgacgaaa tgttgtcttt gcaccgtggt caaggtttgg aattattctg     1620

gagagactct ttgacctgtc catctgaaga agaatacgtc aagatggtct tgggtaagac     1680

cggtggtttg ttcagaattg ctgtcagatt gatgatggcc aagtctgaat gtgacattga     1740

ctttgttcaa ttggttaact tgatttccat ctacttccaa atcagagatg actacatgaa     1800

cttgcaatcc tctgaatacg ctcacaacaa gaacttcgct gaagacttga ctgaaggtaa     1860

gttctccttc ccaaccattc actccattca cgctaaccca tcttccagat tggttatcaa     1920

cactttacaa aagaagtcca cttctccaga aatcttacat cactgtgtca actacatgag     1980

aactgaaacc cactctttcg aatacactca agaagtcttg aacactttat ctggtgcttt     2040

ggaaagagaa ttgggtagat tacaaggtga atttgctgaa gctaactcca agatcgattt     2100

gggtgacgtt gaatctgaag gtagaaccgg taagaacgtc aaattggaag ccatcttgaa     2160

gaaattggct gatatccctc tataaagtga atttacttta aatcttgcat ttaaataaat     2220

tttcttttta tagctttatg acttagtttc aatttatata ctattttaat gacattttcg     2280

attcattgat tgaaagcttt gtgttttttc ttgatgcgct attgcattgt tcttgtcttt     2340

ttcgccacat gtaatatctg tagtagatac ctgatacatt gtggatgctg agtgaaattt     2400

tagttaataa tggaggcgct cttaataatt ttggggatat tggctttttt ttttaaagtt     2460

tacaaatgaa ttttttccgc caggatcctc ttgcccatcg aacgtacaag tactcctctg     2520

ttctctcctt cctttgcttt                                                 2540


<210>  40
<211>  2140
<212>  DNA
<213>  artificial sequence

<220>
<223>  con5 - Strong p (ScFBA1p) - crtE - ScTDH3t - conA expression 
       cassette

<400>  40
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt gtgcctactt       60

ggcttcacat acgttgcata cgtcgatata gataataatg ataatgacag caggattatc      120

gtaatacgta atagttgaaa atctcaaaaa tgtgtgggtc attacgtaaa taatgatagg      180

aatgggattc ttctattttt cctttttcca ttctagcagc cgtcgggaaa acgtggcatc      240

ctctctttcg ggctcaattg gagtcacgct gccgtgagca tcctctcttt ccatatctaa      300

caactgagca cgtaaccaat ggaaaagcat gagcttagcg ttgctccaaa aaagtattgg      360

atggttaata ccatttgtct gttctcttct gactttgact cctcaaaaaa aaaaaatcta      420

caatcaacag atcgcttcaa ttacgccctc acaaaaactt ttttccttct tcttcgccca      480

cgttaaattt tatccctcat gttgtctaac ggatttctgc acttgattta ttataaaaag      540

acaaagacat aatacttctc tatcaatttc agttattgtt cttccttgcg ttattcttct      600

gttcttcttt ttcttttgtc atatataacc ataaccaagt aatacatatt caaaatggac      660

tacgctaaca tcttgactgc cattcctttg gaattcaccc cacaagatga cattgtcttg      720

ttggaaccat accactactt aggtaagaac ccaggtaagg aaatcagatc tcaattgatt      780

gaagctttca actactggtt agatgtcaag aaggaagact tggaagttat ccaaaatgtt      840

gttggtatgt tgcacaccgc ttctttgttg atggatgatg ttgaagattc ttccgtcttg      900

agaagaggtt ctccagttgc tcatttgatc tacggtattc cacaaaccat caacactgct      960

aactacgttt acttcttggc ttaccaagaa atcttcaaat tgcgtccaac tccaattcca     1020

atgccagtta tcccaccatc ttctgcttct ttgcaatctt ctgtctcctc cgcctcctct     1080

tcctcttctg cctcctctga aaacggtggt acctccactc caaactccca aatcccattc     1140

tccaaggaca cctacttgga caaggttatc actgacgaaa tgttgtcttt gcaccgtggt     1200

caaggtttgg aattattctg gagagactct ttgacctgtc catctgaaga agaatacgtc     1260

aagatggtct tgggtaagac cggtggtttg ttcagaattg ctgtcagatt gatgatggcc     1320

aagtctgaat gtgacattga ctttgttcaa ttggttaact tgatttccat ctacttccaa     1380

atcagagatg actacatgaa cttgcaatcc tctgaatacg ctcacaacaa gaacttcgct     1440

gaagacttga ctgaaggtaa gttctccttc ccaaccattc actccattca cgctaaccca     1500

tcttccagat tggttatcaa cactttacaa aagaagtcca cttctccaga aatcttacat     1560

cactgtgtca actacatgag aactgaaacc cactctttcg aatacactca agaagtcttg     1620

aacactttat ctggtgcttt ggaaagagaa ttgggtagat tacaaggtga atttgctgaa     1680

gctaactcca agatcgattt gggtgacgtt gaatctgaag gtagaaccgg taagaacgtc     1740

aaattggaag ccatcttgaa gaaattggct gatatccctc tataaagtga atttacttta     1800

aatcttgcat ttaaataaat tttcttttta tagctttatg acttagtttc aatttatata     1860

ctattttaat gacattttcg attcattgat tgaaagcttt gtgttttttc ttgatgcgct     1920

attgcattgt tcttgtcttt ttcgccacat gtaatatctg tagtagatac ctgatacatt     1980

gtggatgctg agtgaaattt tagttaataa tggaggcgct cttaataatt ttggggatat     2040

tggctttttt ttttaaagtt tacaaatgaa ttttttccgc caggatcctc ttgcccatcg     2100

aacgtacaag tactcctctg ttctctcctt cctttgcttt                           2140


<210>  41
<211>  3431
<212>  DNA
<213>  artificial sequence

<220>
<223>  conA - Low p (KlYDRp) - crtYB - ScPDC1t - conB expression 
       cassette

<400>  41
ttgcccatcg aacgtacaag tactcctctg ttctctcctt cctttgcttt gtgcttttct       60

ttttttgcgg tcacccccat gtggcgggga ggcagaggag taggtagagc aacgaatcct      120

actatttatc caaattagtc taggaactct ttttctagat tttttagatt tgagggcaag      180

cgctgttaac gactcagaaa tgtaagcact acggagtaga acgagaaatc cgccataggt      240

ggaaatccta gcaaaatctt gcttacccta gctagcctca ggtaagctag ccttagcctg      300

tcaaattttt ttcaaaattt ggtaagtttc tactagcaaa gcaaacacgg ttcaacaaac      360

cgaaaactcc actcattata cgtggaaacc gaaacaaaaa aacaaaaacc aaaatactcg      420

ccaatgagaa agttgctgcg tttctacttt cgaggaagag gaactgagag gattgactac      480

gaaaggggca aaaacgagtc gtattctccc attattgtct gctaccacgc ggtctagtag      540

aataagcaac cagtcaacgc taagacaggt aatcaaaata ccagtctgct ggctacgggc      600

tagtttttac ctcttttaga acccactgta aaagtccgtt gtaaagcccg ttctcactgt      660

tggcgttttt ttttttttgg tttagtttct tatttttcat ttttttcttt catgaccaaa      720

aacaaacaaa tctcgcgatt tgtactgcgg ccactggggc gtggccaaaa aaatgacaaa      780

tttagaaacc ttagtttctg atttttcctg ttatgaggag atatgataaa aaatattact      840

gctttattgt ttttttttta tctactgaaa tagagaaact tacccaagga ggaggcaaaa      900

aaaagagtat atatacagca ggtaccattc agattttaat atattctttt ctcttcttct      960

acactattat tataataatt ttactatatt catttttagc ttaaaacctc atagaatatt     1020

attcttcagt cactcgctta aatacttatc aaaaatgacc gctttggctt actaccaaat     1080

ccacttgatc tacactttgc caatcttagg tttgctaggt ttgttgactt ctccaatttt     1140

gaccaaattc gacatctaca agatttctat cttagtcttt attgctttct ctgctaccac     1200

tccatgggac tcctggatca tcagaaacgg tgcctggacc tacccatctg ctgaatctgg     1260

tcaaggtgtt ttcggtacct ttttggatgt cccatacgaa gaatacgcct tctttgttat     1320

ccaaaccgtc atcaccggtt tggtttacgt tttggctacc agacatttgt tgccatcttt     1380

ggctctacca aagacccgtt cttctgcctt gtctctagct ttgaaggctt taatcccatt     1440

gccaatcatc tatttgttca ccgctcatcc atctccatcc ccagatcctt tggttactga     1500

ccactacttc tacatgagag ctttgtcttt gttgatcacc ccaccaacca tgttgttggc     1560

tgctttatct ggtgaatacg ctttcgactg gaaatctggt agagctaagt ccaccattgc     1620

tgccatcatg atcccaactg tctacttgat ctgggttgac tacgttgccg ttggtcaaga     1680

ctcctggtcc atcaacgatg aaaagattgt cggttggaga ttaggtggtg tcttgccaat     1740

tgaagaagct atgttcttct tattgaccaa cttgatgatc gttttgggtt tgtctgcctg     1800

tgaccacact caagccttgt acttgttgca cggtagaact atctacggta acaagaagat     1860

gccatcttct ttcccattaa tcactccacc agttttgtcc ttgttcttct cctccagacc     1920

atactcctcc caaccaaaga gagatttgga attggctgtc aagttgttgg aaaagaagtc     1980

cagatctttc ttcgttgctt ctgccggttt cccatctgaa gtcagagaaa gattggttgg     2040

tttgtacgct ttctgtcgtg tcaccgatga cttgattgac tctccagaag tttcctccaa     2100

cccacacgct accattgaca tggtttccga tttcttgact ttattattcg gtcctccatt     2160

gcacccatct caaccagaca agattttgtc ttctccatta ttaccacctt cccacccatc     2220

cagaccaact ggtatgtacc cattaccacc acctccatct ttgtctccag ctgaattggt     2280

ccaattcttg actgaacgtg tcccagttca ataccacttc gctttcagat tgttggccaa     2340

attgcaaggt ttgattccaa gatacccatt ggatgaatta ttgagaggtt acaccactga     2400

cttgatcttc ccattgtcca ctgaagccgt ccaagctaga aagaccccaa ttgaaactac     2460

tgctgacttg ttggactacg gtttgtgtgt tgccggttct gttgctgaat tgttggtcta     2520

cgtttcctgg gcttccgctc catcccaagt tccagctact attgaagaaa gagaagctgt     2580

tttggtcgcc tctcgtgaaa tgggtaccgc tttgcaattg gtcaacattg ccagagatat     2640

caagggtgac gctactgaag gtagattcta cttgccattg tctttctttg gtttgagaga     2700

tgaatccaaa ttggccattc caactgactg gactgaacca agacctcaag atttcgacaa     2760

attgttgtct ctatctccat cttccacttt accatcctct aacgcttctg aatccttcag     2820

attcgaatgg aagacctact ctttgccatt ggttgcttac gctgaagatt tggctaagca     2880

ctcttacaag ggtattgaca gattaccaac tgaagtccaa gctggtatga gagctgcttg     2940

tgcttcttac ttgttgattg gtcgtgaaat caaggttgtc tggaagggtg atgtcggtga     3000

aagaagaacc gttgctggtt ggagaagagt cagaaaggtt ttgtctgttg tcatgtccgg     3060

ttgggaaggt caataaagcg atttaatctc taattattag ttaaagtttt ataagcattt     3120

ttatgtaacg aaaaataaat tggttcatat tattactgca ctgtcactta ccatggaaag     3180

accagacaag aagttgccga cagtctgttg aattggcctg gttaggctta agtctgggtc     3240

cgcttcttta caaatttgga gaatttctct taaacgatat gtatattctt ttcgttggaa     3300

aagatgtctt ccaaaaaaaa aaccgatgaa ttagtggaac caaggaaaaa aaaagaggta     3360

tccttgatta aggaacacct ccggatcgat gtacacaacc gactgcaccc aaacgaacac     3420

aaatcttagc a                                                          3431


<210>  42
<211>  3431
<212>  DNA
<213>  artificial sequence

<220>
<223>  conA - Med p (KlTEF2p) - crtYB - ScPDC1t - conB expression 
       cassette

<400>  42
ttgcccatcg aacgtacaag tactcctctg ttctctcctt cctttgcttt gtgcgagcct       60

gtccaagcaa atgccttctc ataaatggtg ccaaagaccc gcaagcccaa agcaattacc      120

ccccaaaaag aaatgatata gtgcaagata cgtatatgac catgacttga ctaggtgaaa      180

cagtgcagaa acagccgcac aaaagcagcc ctaaccctca gagtcgattt tactctttca      240

ggtaataaag cctcgacatc aattttagac agaagccagg ctggcctcga gattatagcc      300

ataggcaagc aagaggagag aaggggaggc cccccatggg gggcctcccc cccgctgtca      360

aggtttggca gaacctagct tcattaggcc actagcccag cctaaaacgt caacgggcag      420

gaggaacact cccacaagac ggcgtagtat tctcgattca taaccatttt ctcaatcgaa      480

ttacacagaa cacaccgtac aaacctctct atcataacta cttaatagtc acacacgtac      540

tcgtctaaat acacatcatc gtcctacaag ttcatcaaag tgttggacag acaactatac      600

cagcatggat ctcttgtatc ggttcttttc tcccgctctc tcgcaataac aatgaacact      660

gggtcaatca tagcctacac aggtgaacag agtagcgttt atacagggtt tatacggtga      720

ttcctacggc aaaaattttt catttctaaa aaaaaaaaga aaaatttttc tttccaacgc      780

tagaaggaaa agaaaaatct aattaaattg atttggtgat tttctgagag ttcccttttt      840

catatatcga attttgaata taaaaggaga tcgaaaaaat ttttctattc aatctgtttt      900

ctggttttat ttgatagttt ttttgtgtat tattattatg gattagtact ggtttatatg      960

ggtttttctg tataacttct ttttatttta gtttgtttaa tcttattttg agttacatta     1020

tagttcccta actgcaagag aagtaacatt aaaaatgacc gctttggctt actaccaaat     1080

ccacttgatc tacactttgc caatcttagg tttgctaggt ttgttgactt ctccaatttt     1140

gaccaaattc gacatctaca agatttctat cttagtcttt attgctttct ctgctaccac     1200

tccatgggac tcctggatca tcagaaacgg tgcctggacc tacccatctg ctgaatctgg     1260

tcaaggtgtt ttcggtacct ttttggatgt cccatacgaa gaatacgcct tctttgttat     1320

ccaaaccgtc atcaccggtt tggtttacgt tttggctacc agacatttgt tgccatcttt     1380

ggctctacca aagacccgtt cttctgcctt gtctctagct ttgaaggctt taatcccatt     1440

gccaatcatc tatttgttca ccgctcatcc atctccatcc ccagatcctt tggttactga     1500

ccactacttc tacatgagag ctttgtcttt gttgatcacc ccaccaacca tgttgttggc     1560

tgctttatct ggtgaatacg ctttcgactg gaaatctggt agagctaagt ccaccattgc     1620

tgccatcatg atcccaactg tctacttgat ctgggttgac tacgttgccg ttggtcaaga     1680

ctcctggtcc atcaacgatg aaaagattgt cggttggaga ttaggtggtg tcttgccaat     1740

tgaagaagct atgttcttct tattgaccaa cttgatgatc gttttgggtt tgtctgcctg     1800

tgaccacact caagccttgt acttgttgca cggtagaact atctacggta acaagaagat     1860

gccatcttct ttcccattaa tcactccacc agttttgtcc ttgttcttct cctccagacc     1920

atactcctcc caaccaaaga gagatttgga attggctgtc aagttgttgg aaaagaagtc     1980

cagatctttc ttcgttgctt ctgccggttt cccatctgaa gtcagagaaa gattggttgg     2040

tttgtacgct ttctgtcgtg tcaccgatga cttgattgac tctccagaag tttcctccaa     2100

cccacacgct accattgaca tggtttccga tttcttgact ttattattcg gtcctccatt     2160

gcacccatct caaccagaca agattttgtc ttctccatta ttaccacctt cccacccatc     2220

cagaccaact ggtatgtacc cattaccacc acctccatct ttgtctccag ctgaattggt     2280

ccaattcttg actgaacgtg tcccagttca ataccacttc gctttcagat tgttggccaa     2340

attgcaaggt ttgattccaa gatacccatt ggatgaatta ttgagaggtt acaccactga     2400

cttgatcttc ccattgtcca ctgaagccgt ccaagctaga aagaccccaa ttgaaactac     2460

tgctgacttg ttggactacg gtttgtgtgt tgccggttct gttgctgaat tgttggtcta     2520

cgtttcctgg gcttccgctc catcccaagt tccagctact attgaagaaa gagaagctgt     2580

tttggtcgcc tctcgtgaaa tgggtaccgc tttgcaattg gtcaacattg ccagagatat     2640

caagggtgac gctactgaag gtagattcta cttgccattg tctttctttg gtttgagaga     2700

tgaatccaaa ttggccattc caactgactg gactgaacca agacctcaag atttcgacaa     2760

attgttgtct ctatctccat cttccacttt accatcctct aacgcttctg aatccttcag     2820

attcgaatgg aagacctact ctttgccatt ggttgcttac gctgaagatt tggctaagca     2880

ctcttacaag ggtattgaca gattaccaac tgaagtccaa gctggtatga gagctgcttg     2940

tgcttcttac ttgttgattg gtcgtgaaat caaggttgtc tggaagggtg atgtcggtga     3000

aagaagaacc gttgctggtt ggagaagagt cagaaaggtt ttgtctgttg tcatgtccgg     3060

ttgggaaggt caataaagcg atttaatctc taattattag ttaaagtttt ataagcattt     3120

ttatgtaacg aaaaataaat tggttcatat tattactgca ctgtcactta ccatggaaag     3180

accagacaag aagttgccga cagtctgttg aattggcctg gttaggctta agtctgggtc     3240

cgcttcttta caaatttgga gaatttctct taaacgatat gtatattctt ttcgttggaa     3300

aagatgtctt ccaaaaaaaa aaccgatgaa ttagtggaac caaggaaaaa aaaagaggta     3360

tccttgatta aggaacacct ccggatcgat gtacacaacc gactgcaccc aaacgaacac     3420

aaatcttagc a                                                          3431


<210>  43
<211>  3031
<212>  DNA
<213>  artificial sequence

<220>
<223>  conA - Strong p (ScTEF1p) - crtYB - ScPDC1t - conB expression 
       cassette

<400>  43
ttgcccatcg aacgtacaag tactcctctg ttctctcctt cctttgcttt gtgcttggct       60

gataatagcg tataaacaat gcatactttg tacgttcaaa atacaatgca gtagatatat      120

ttatgcatat tacatataat acatatcaca taggaagcaa caggcgcgtt ggacttttaa      180

ttttcgagga ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat cccccacaca      240

ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg      300

catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc      360

tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga caccgcctcg      420

tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa      480

attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa      540

cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac      600

ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaatgacc      660

gctttggctt actaccaaat ccacttgatc tacactttgc caatcttagg tttgctaggt      720

ttgttgactt ctccaatttt gaccaaattc gacatctaca agatttctat cttagtcttt      780

attgctttct ctgctaccac tccatgggac tcctggatca tcagaaacgg tgcctggacc      840

tacccatctg ctgaatctgg tcaaggtgtt ttcggtacct ttttggatgt cccatacgaa      900

gaatacgcct tctttgttat ccaaaccgtc atcaccggtt tggtttacgt tttggctacc      960

agacatttgt tgccatcttt ggctctacca aagacccgtt cttctgcctt gtctctagct     1020

ttgaaggctt taatcccatt gccaatcatc tatttgttca ccgctcatcc atctccatcc     1080

ccagatcctt tggttactga ccactacttc tacatgagag ctttgtcttt gttgatcacc     1140

ccaccaacca tgttgttggc tgctttatct ggtgaatacg ctttcgactg gaaatctggt     1200

agagctaagt ccaccattgc tgccatcatg atcccaactg tctacttgat ctgggttgac     1260

tacgttgccg ttggtcaaga ctcctggtcc atcaacgatg aaaagattgt cggttggaga     1320

ttaggtggtg tcttgccaat tgaagaagct atgttcttct tattgaccaa cttgatgatc     1380

gttttgggtt tgtctgcctg tgaccacact caagccttgt acttgttgca cggtagaact     1440

atctacggta acaagaagat gccatcttct ttcccattaa tcactccacc agttttgtcc     1500

ttgttcttct cctccagacc atactcctcc caaccaaaga gagatttgga attggctgtc     1560

aagttgttgg aaaagaagtc cagatctttc ttcgttgctt ctgccggttt cccatctgaa     1620

gtcagagaaa gattggttgg tttgtacgct ttctgtcgtg tcaccgatga cttgattgac     1680

tctccagaag tttcctccaa cccacacgct accattgaca tggtttccga tttcttgact     1740

ttattattcg gtcctccatt gcacccatct caaccagaca agattttgtc ttctccatta     1800

ttaccacctt cccacccatc cagaccaact ggtatgtacc cattaccacc acctccatct     1860

ttgtctccag ctgaattggt ccaattcttg actgaacgtg tcccagttca ataccacttc     1920

gctttcagat tgttggccaa attgcaaggt ttgattccaa gatacccatt ggatgaatta     1980

ttgagaggtt acaccactga cttgatcttc ccattgtcca ctgaagccgt ccaagctaga     2040

aagaccccaa ttgaaactac tgctgacttg ttggactacg gtttgtgtgt tgccggttct     2100

gttgctgaat tgttggtcta cgtttcctgg gcttccgctc catcccaagt tccagctact     2160

attgaagaaa gagaagctgt tttggtcgcc tctcgtgaaa tgggtaccgc tttgcaattg     2220

gtcaacattg ccagagatat caagggtgac gctactgaag gtagattcta cttgccattg     2280

tctttctttg gtttgagaga tgaatccaaa ttggccattc caactgactg gactgaacca     2340

agacctcaag atttcgacaa attgttgtct ctatctccat cttccacttt accatcctct     2400

aacgcttctg aatccttcag attcgaatgg aagacctact ctttgccatt ggttgcttac     2460

gctgaagatt tggctaagca ctcttacaag ggtattgaca gattaccaac tgaagtccaa     2520

gctggtatga gagctgcttg tgcttcttac ttgttgattg gtcgtgaaat caaggttgtc     2580

tggaagggtg atgtcggtga aagaagaacc gttgctggtt ggagaagagt cagaaaggtt     2640

ttgtctgttg tcatgtccgg ttgggaaggt caataaagcg atttaatctc taattattag     2700

ttaaagtttt ataagcattt ttatgtaacg aaaaataaat tggttcatat tattactgca     2760

ctgtcactta ccatggaaag accagacaag aagttgccga cagtctgttg aattggcctg     2820

gttaggctta agtctgggtc cgcttcttta caaatttgga gaatttctct taaacgatat     2880

gtatattctt ttcgttggaa aagatgtctt ccaaaaaaaa aaccgatgaa ttagtggaac     2940

caaggaaaaa aaaagaggta tccttgatta aggaacacct ccggatcgat gtacacaacc     3000

gactgcaccc aaacgaacac aaatcttagc a                                    3031


<210>  44
<211>  2758
<212>  DNA
<213>  artificial sequence

<220>
<223>  conB - Low p (ScPRE3p) - crtI - ScTAL1t - conC expression 
       cassette

<400>  44
cggatcgatg tacacaaccg actgcaccca aacgaacaca aatcttagca gtgccaaaca       60

ttaatttgtt ctgcatactt tgaacctttc agaaaataaa aaacattacg cgcatactta      120

ccctgctcgc gaagaagagt aacactaacg cattctatgg gcaattgaag acagtattca      180

gtacaagaca tagtccgttt ccttgagtca attcctatag cattatgaac tagccgcctt      240

taagagtgcc aagctgttca acaccgatca tttttgatga tttggcgttt ttgttatatt      300

gatagatttc ttttgaattt tgtcattttc acttttccac tcgcaacgga atccggtggc      360

aaaaaaggga aaagcattga aatgcaatct ttaacagtat tttaaacaag ttgcgacacg      420

gtgtacaatt acgataagaa ttgctacttc aaagtacaca cagaaagtta acatgaatgg      480

aattcaagtg gacatcaatc gtttgaaaaa gggcgaagtc agtttaggta cctcaatgta      540

tgtatataag aatttttcct cccactttat tgtttctaaa agttcaatga agtaaagtct      600

caattggcct tattactaac taataggtat cttataatca cctaataaaa tagaatgggt      660

aaggaacaag accaagacaa gccaactgcc atcatcgttg gttgtggtat cggtggtatt      720

gctaccgctg ccagattagc taaggaaggt ttccaagtta ccgtctttga aaagaacgac      780

tactccggtg gtagatgttc tttgattgaa agagatggtt acagattcga ccaaggtcca      840

tctttgttgc tattaccaga cttgttcaag caaaccttcg aagatttggg tgaaaagatg      900

gaagactggg ttgatttgat caagtgtgaa ccaaactacg tttgtcactt ccatgatgaa      960

gaaactttca ccttctccac tgacatggct ttattgaaga gagaagtcga aagatttgaa     1020

ggtaaagatg gtttcgacag attcttgtct ttcatccaag aagctcacag acattacgaa     1080

ttggctgttg tccacgtctt gcaaaagaac ttcccaggtt tcgctgcttt cttgagatta     1140

caattcatcg gtcaaatctt agctttgcac ccatttgaat ccatctggac cagagtttgt     1200

cgttacttca agactgacag attgagaaga gtcttctcct ttgccgttat gtacatgggt     1260

caatctccat actctgctcc aggtacctac tccttgttgc aatacactga attgactgaa     1320

ggtatctggt acccaagagg tggtttctgg caagttccaa acactttgtt gcaaatcgtc     1380

aagagaaaca acccatctgc taagttcaac ttcaacgctc cagtttctca agttttgttg     1440

tctccagcta aggacagagc taccggtgtc agattagaat ctggtgaaga acaccacgct     1500

gatgttgtca ttgtcaatgc tgacttggtc tacgcttctg aacatttgat tccagatgat     1560

gctagaaaca agatcggtca attaggtgaa gttaagcgtt cctggtgggc tgatttggtt     1620

ggtggtaaga agttgaaggg ttcttgttct tctttgtctt tctactggtc tatggacaga     1680

atcgttgacg gtttgggtgg tcacaacatc ttcttggctg aagacttcaa gggttccttc     1740

gacaccattt tcgaagaatt gggtttgcca gctgacccat ctttctatgt taacgttcca     1800

tccagaattg acccttctgc tgctccagaa ggtaaggatg ccattgtcat cttagtccca     1860

tgtggtcaca tcgatgcttc caaccctcaa gactacaaca aattggttgc cagagccaga     1920

aagttcgtca tccaaacctt gtctgccaag ttgggtctac cagatttcga aaagatgatt     1980

gttgctgaaa aggttcacga tgctccatcc tgggaaaagg aattcaactt gaaggacggt     2040

tccattttgg gtttggctca caacttcatg caagtcttgg gtttcagacc atccaccaga     2100

cacccaaagt acgacaaatt gttctttgtc ggtgcttcta cccacccagg tactggtgtt     2160

ccaattgtct tggctggtgc caaattgact gctaaccaag ttttggaatc cttcgatcgt     2220

tctccagctc cagatcctaa catgtctttg tctgttccat acggtaagcc attgaaatcc     2280

aacggtactg gtattgactc tcaagtccaa ttgaaattca tggacttgga acgttgggtt     2340

tacctattag tcttgttgat tggtgctgtt atcgccagat ccgtcggtgt cttggccttt     2400

taaaggaagt atctcggaaa tattaattta ggccatgtcc ttatgcacgt ttcttttgat     2460

acttacgggt acatgtacac aagtatatct atatatataa attaatgaaa atcccctatt     2520

tatatatatg actttaacga gacagaacag ttttttattt tttatcctat ttgatgaatg     2580

atacagtttc ttattcacgt gttataccca caccaaatcc aatagcaata ccggccatca     2640

caatcactgt ttcggcagcc cctaagatca gacaaaacat ccggaaccac cttaaatcaa     2700

cgtccctcac gctttccggc atcttccaga ccacagtata tccatccgcc tcctgttg       2758


<210>  45
<211>  2758
<212>  DNA
<213>  artificial sequence

<220>
<223>  conB - Med p (ScACT1p) - crtI - ScTAL1t - conC expression 
       cassette

<400>  45
cggatcgatg tacacaaccg actgcaccca aacgaacaca aatcttagca gtgcaacata       60

tatacacaat tacagtaaca ataacaagag gacagatact accaaaatgt gtggggaagc      120

gggtaagctg ccacagcaat taatgcacaa catttaacct acattcttcc ttatcggatc      180

ctcaaaaccc ttaaaaacat atgcctcacc ctaacatatt ttccaattaa ccctcaatat      240

ttctctgtca cccggcctct attttccatt ttcttcttta cccgccacgc gtttttttct      300

ttcaaatttt tttcttcttt cttctttttc ttccacgtcc tcttgcataa ataaataaac      360

cgttttgaaa ccaaactcgc ctctctctct cctttttgaa atatttttgg gtttgtttga      420

tcctttcctt cccaatctct cttgtttaat atatattcat ttatatcacg ctctcttttt      480

atcttccttt ttttcctctc tcttgtattc ttccttcccc tttctactca aaccaagaag      540

aaaaagaaaa ggtcaatctt tgttaaagaa taggatcttc tactacatca gcttttagat      600

ttttcacgct tactgctttt ttcttcccaa gatcgaaaat ttactgaatt aacaatgggt      660

aaggaacaag accaagacaa gccaactgcc atcatcgttg gttgtggtat cggtggtatt      720

gctaccgctg ccagattagc taaggaaggt ttccaagtta ccgtctttga aaagaacgac      780

tactccggtg gtagatgttc tttgattgaa agagatggtt acagattcga ccaaggtcca      840

tctttgttgc tattaccaga cttgttcaag caaaccttcg aagatttggg tgaaaagatg      900

gaagactggg ttgatttgat caagtgtgaa ccaaactacg tttgtcactt ccatgatgaa      960

gaaactttca ccttctccac tgacatggct ttattgaaga gagaagtcga aagatttgaa     1020

ggtaaagatg gtttcgacag attcttgtct ttcatccaag aagctcacag acattacgaa     1080

ttggctgttg tccacgtctt gcaaaagaac ttcccaggtt tcgctgcttt cttgagatta     1140

caattcatcg gtcaaatctt agctttgcac ccatttgaat ccatctggac cagagtttgt     1200

cgttacttca agactgacag attgagaaga gtcttctcct ttgccgttat gtacatgggt     1260

caatctccat actctgctcc aggtacctac tccttgttgc aatacactga attgactgaa     1320

ggtatctggt acccaagagg tggtttctgg caagttccaa acactttgtt gcaaatcgtc     1380

aagagaaaca acccatctgc taagttcaac ttcaacgctc cagtttctca agttttgttg     1440

tctccagcta aggacagagc taccggtgtc agattagaat ctggtgaaga acaccacgct     1500

gatgttgtca ttgtcaatgc tgacttggtc tacgcttctg aacatttgat tccagatgat     1560

gctagaaaca agatcggtca attaggtgaa gttaagcgtt cctggtgggc tgatttggtt     1620

ggtggtaaga agttgaaggg ttcttgttct tctttgtctt tctactggtc tatggacaga     1680

atcgttgacg gtttgggtgg tcacaacatc ttcttggctg aagacttcaa gggttccttc     1740

gacaccattt tcgaagaatt gggtttgcca gctgacccat ctttctatgt taacgttcca     1800

tccagaattg acccttctgc tgctccagaa ggtaaggatg ccattgtcat cttagtccca     1860

tgtggtcaca tcgatgcttc caaccctcaa gactacaaca aattggttgc cagagccaga     1920

aagttcgtca tccaaacctt gtctgccaag ttgggtctac cagatttcga aaagatgatt     1980

gttgctgaaa aggttcacga tgctccatcc tgggaaaagg aattcaactt gaaggacggt     2040

tccattttgg gtttggctca caacttcatg caagtcttgg gtttcagacc atccaccaga     2100

cacccaaagt acgacaaatt gttctttgtc ggtgcttcta cccacccagg tactggtgtt     2160

ccaattgtct tggctggtgc caaattgact gctaaccaag ttttggaatc cttcgatcgt     2220

tctccagctc cagatcctaa catgtctttg tctgttccat acggtaagcc attgaaatcc     2280

aacggtactg gtattgactc tcaagtccaa ttgaaattca tggacttgga acgttgggtt     2340

tacctattag tcttgttgat tggtgctgtt atcgccagat ccgtcggtgt cttggccttt     2400

taaaggaagt atctcggaaa tattaattta ggccatgtcc ttatgcacgt ttcttttgat     2460

acttacgggt acatgtacac aagtatatct atatatataa attaatgaaa atcccctatt     2520

tatatatatg actttaacga gacagaacag ttttttattt tttatcctat ttgatgaatg     2580

atacagtttc ttattcacgt gttataccca caccaaatcc aatagcaata ccggccatca     2640

caatcactgt ttcggcagcc cctaagatca gacaaaacat ccggaaccac cttaaatcaa     2700

cgtccctcac gctttccggc atcttccaga ccacagtata tccatccgcc tcctgttg       2758


<210>  46
<211>  3158
<212>  DNA
<213>  artificial sequence

<220>
<223>  conB - Strong p (KlENO1p) - crtI - ScTAL1t - conC expression 
       cassette

<400>  46
cggatcgatg tacacaaccg actgcaccca aacgaacaca aatcttagca gtgccgtatc       60

cctatctgga ttaacatcac tgccacagat cgaattgcaa gaagccacac ttcacgtgat      120

ccactcgttc atcaggtttg tagcttcatg gcgcaggact tctgatgatg aactatctgg      180

ctcatccgga tggatcacaa ggatggatac cctcagacag tacgtttccg tatggagcga      240

tcttacaaga accaacagtt gatcctatta cttttttttt attttttgtc cctccgggat      300

ggcaagaggg acaaagaaga atcttcgttc ttctttcttg ttctcaactt cccagcttcc      360

gtgtgattac cctccgggac aacagaaaaa ctggcattcg gtatcccggg aatctgctga      420

gaaggaaaga aaacgaaaaa aaaattgtac atttgtgtca cattatgaat tacaggaagt      480

cagaaaacag gcagcacatg tctcgcacat gcatgtccat cagacgagac attatgagac      540

atgcacgcgt gtgagagaca tagcaaaagt ctctccagta cacacagaaa gacacgttca      600

caatccaggc accccacaga gaaaaaaaaa agaagaagcc cggaagctgg cacgccatca      660

tcaaccaccg ctcggtttac acgcatccca actgtctttt ttttctggaa tcctataata      720

actggcatct ggaaatcacg ttgtatgttg caccatagtg actggctgtc tgactagcaa      780

acattgattc cctgattccc atttggctca attttgatga gaaacagttg attgattctt      840

gtcaattttt ttttctttgg accaccacca accaattgac attgaagtac tttcccatga      900

tttgaggtta tataaaagga cgttcaaatc actttcaagg ttaattcagt tttgtcaatt      960

gatttaagtt caattgttaa caaatttaat ttaattcgaa acaaaccaaa ccaattcatt     1020

tgaattaaca aaccaaccca caaaacaaaa aaaaatgggt aaggaacaag accaagacaa     1080

gccaactgcc atcatcgttg gttgtggtat cggtggtatt gctaccgctg ccagattagc     1140

taaggaaggt ttccaagtta ccgtctttga aaagaacgac tactccggtg gtagatgttc     1200

tttgattgaa agagatggtt acagattcga ccaaggtcca tctttgttgc tattaccaga     1260

cttgttcaag caaaccttcg aagatttggg tgaaaagatg gaagactggg ttgatttgat     1320

caagtgtgaa ccaaactacg tttgtcactt ccatgatgaa gaaactttca ccttctccac     1380

tgacatggct ttattgaaga gagaagtcga aagatttgaa ggtaaagatg gtttcgacag     1440

attcttgtct ttcatccaag aagctcacag acattacgaa ttggctgttg tccacgtctt     1500

gcaaaagaac ttcccaggtt tcgctgcttt cttgagatta caattcatcg gtcaaatctt     1560

agctttgcac ccatttgaat ccatctggac cagagtttgt cgttacttca agactgacag     1620

attgagaaga gtcttctcct ttgccgttat gtacatgggt caatctccat actctgctcc     1680

aggtacctac tccttgttgc aatacactga attgactgaa ggtatctggt acccaagagg     1740

tggtttctgg caagttccaa acactttgtt gcaaatcgtc aagagaaaca acccatctgc     1800

taagttcaac ttcaacgctc cagtttctca agttttgttg tctccagcta aggacagagc     1860

taccggtgtc agattagaat ctggtgaaga acaccacgct gatgttgtca ttgtcaatgc     1920

tgacttggtc tacgcttctg aacatttgat tccagatgat gctagaaaca agatcggtca     1980

attaggtgaa gttaagcgtt cctggtgggc tgatttggtt ggtggtaaga agttgaaggg     2040

ttcttgttct tctttgtctt tctactggtc tatggacaga atcgttgacg gtttgggtgg     2100

tcacaacatc ttcttggctg aagacttcaa gggttccttc gacaccattt tcgaagaatt     2160

gggtttgcca gctgacccat ctttctatgt taacgttcca tccagaattg acccttctgc     2220

tgctccagaa ggtaaggatg ccattgtcat cttagtccca tgtggtcaca tcgatgcttc     2280

caaccctcaa gactacaaca aattggttgc cagagccaga aagttcgtca tccaaacctt     2340

gtctgccaag ttgggtctac cagatttcga aaagatgatt gttgctgaaa aggttcacga     2400

tgctccatcc tgggaaaagg aattcaactt gaaggacggt tccattttgg gtttggctca     2460

caacttcatg caagtcttgg gtttcagacc atccaccaga cacccaaagt acgacaaatt     2520

gttctttgtc ggtgcttcta cccacccagg tactggtgtt ccaattgtct tggctggtgc     2580

caaattgact gctaaccaag ttttggaatc cttcgatcgt tctccagctc cagatcctaa     2640

catgtctttg tctgttccat acggtaagcc attgaaatcc aacggtactg gtattgactc     2700

tcaagtccaa ttgaaattca tggacttgga acgttgggtt tacctattag tcttgttgat     2760

tggtgctgtt atcgccagat ccgtcggtgt cttggccttt taaaggaagt atctcggaaa     2820

tattaattta ggccatgtcc ttatgcacgt ttcttttgat acttacgggt acatgtacac     2880

aagtatatct atatatataa attaatgaaa atcccctatt tatatatatg actttaacga     2940

gacagaacag ttttttattt tttatcctat ttgatgaatg atacagtttc ttattcacgt     3000

gttataccca caccaaatcc aatagcaata ccggccatca caatcactgt ttcggcagcc     3060

cctaagatca gacaaaacat ccggaaccac cttaaatcaa cgtccctcac gctttccggc     3120

atcttccaga ccacagtata tccatccgcc tcctgttg                             3158


<210>  47
<211>  2534
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT1 - Low p (KlTDH2p) - crtE - 
       ScTDH3t - Homology to INT1

<400>  47
catgcctatc cggcggcaac attattactc tggtatacgg gcgtaactcc acgtaaaaac       60

taaaacgagc ccccaccaaa gaacaaaaaa gaaggtgctg ggcccccact ttcttccctt      120

gcacgtgata ggaagatggc tacagaaaca agaagatgga aatcgaagga aagagggaga      180

ctggaagctg taaaaactga aatgaaaaaa aaaaaaaaaa aaaaaaaaca agaagctgaa      240

aatggaagac tgaaatttga aaaatggtaa aaaaaaaaaa gaaacacgaa gctaaaaacc      300

tggattccat tttgagaaga agcaagaaag gtaagtatgg taacgaccgt acaggcaagc      360

gcgaaggcaa atggaaaagc tggagtccgg aagataatca tttcatcttc ttttgttaga      420

acagaacagt ggatgtccct catctcggta acgtattgtc catgccctag aactctctgt      480

ccctaaaaag aggacaaaaa cccaatggtt tccccagctt ccagtggagc caccgatccc      540

actggaaacc actggacagg aagagaaaat cacggacttc ctctattgaa ggataattca      600

acactttcac cagatcccaa atgtcccgcc cctattcccg tgttccatca cgtaccataa      660

cttaccattt catcacgttc tctatggcac actggtactg cttcgactgc tttgcttcat      720

cttctctatg ggccaatgag ctaatgagca caatgtgctg cgaaataaag ggatatctaa      780

tttatattat tacattataa tatgtactag tgtggttatt ggtaattgta cttaattttg      840

atatataaag ggtggatctt tttcattttg aatcagaatt ggaattgcaa cttgtctctt      900

gtcactatta cttaatagta attatatttc ttattaacct tttttttaag tcaaaacacc      960

aaggacaaga actactcttc aaaggtattt caagttatca tacgtgtcac acacgcttca     1020

cagtttcaag taaaaaaaaa gaatattaca caatggacta cgctaacatc ttgactgcca     1080

ttcctttgga attcacccca caagatgaca ttgtcttgtt ggaaccatac cactacttag     1140

gtaagaaccc aggtaaggaa atcagatctc aattgattga agctttcaac tactggttag     1200

atgtcaagaa ggaagacttg gaagttatcc aaaatgttgt tggtatgttg cacaccgctt     1260

ctttgttgat ggatgatgtt gaagattctt ccgtcttgag aagaggttct ccagttgctc     1320

atttgatcta cggtattcca caaaccatca acactgctaa ctacgtttac ttcttggctt     1380

accaagaaat cttcaaattg cgtccaactc caattccaat gccagttatc ccaccatctt     1440

ctgcttcttt gcaatcttct gtctcctccg cctcctcttc ctcttctgcc tcctctgaaa     1500

acggtggtac ctccactcca aactcccaaa tcccattctc caaggacacc tacttggaca     1560

aggttatcac tgacgaaatg ttgtctttgc accgtggtca aggtttggaa ttattctgga     1620

gagactcttt gacctgtcca tctgaagaag aatacgtcaa gatggtcttg ggtaagaccg     1680

gtggtttgtt cagaattgct gtcagattga tgatggccaa gtctgaatgt gacattgact     1740

ttgttcaatt ggttaacttg atttccatct acttccaaat cagagatgac tacatgaact     1800

tgcaatcctc tgaatacgct cacaacaaga acttcgctga agacttgact gaaggtaagt     1860

tctccttccc aaccattcac tccattcacg ctaacccatc ttccagattg gttatcaaca     1920

ctttacaaaa gaagtccact tctccagaaa tcttacatca ctgtgtcaac tacatgagaa     1980

ctgaaaccca ctctttcgaa tacactcaag aagtcttgaa cactttatct ggtgctttgg     2040

aaagagaatt gggtagatta caaggtgaat ttgctgaagc taactccaag atcgatttgg     2100

gtgacgttga atctgaaggt agaaccggta agaacgtcaa attggaagcc atcttgaaga     2160

aattggctga tatccctcta taaagtgaat ttactttaaa tcttgcattt aaataaattt     2220

tctttttata gctttatgac ttagtttcaa tttatatact attttaatga cattttcgat     2280

tcattgattg aaagctttgt gttttttctt gatgcgctat tgcattgttc ttgtcttttt     2340

cgccacatgt aatatctgta gtagatacct gatacattgt ggatgctgag tgaaatttta     2400

gttaataatg gaggcgctct taataatttt ggggatattg gctttttttt ttaaagttta     2460

caaatgaatt ttttccgcca ggatgaagta tggatctttg aaaccctttt attttatttt     2520

acttttttta gaat                                                       2534


<210>  48
<211>  2530
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT1 - Med p (KlPGK1p) - crtE - 
       ScTDH3t - Homology to INT1

<400>  48
catgcctatc cggcggcaac attattactc tggtatacgg gcgtaactcc acctcatcac       60

tagaagccga actgttgtct tcagtgggga ttggttcgac attttgccaa ttgctgtcga      120

tgtacccttt caaagccatg taccttaaat cttcatcctt ggcaagtaga ttcatcgggt      180

gtgtttgaag taagaatatt tgcttgtttt tatggtatca aaggtatatg ttgtagaaga      240

caatttccgg taatccaatt gtctgtctgc tcagtttagc acatgtatag tacgttgcac      300

atagtctaca atattcagca ttcagcattc agtatacagc atatggctaa atgatcacaa      360

atgtgattga tgatttgaca cgactagaaa agagaacgaa aaagggaaat tccatgtcac      420

gtgcgttggc acgtgacatg gaatatcgaa gaaagaaaaa aaaaacgatc tcgtcctagt      480

ggaagcccag agtctggtcc ccccggagtc ttcccaaaac aagaagctga cacatgttga      540

cacagaacac cccacagcaa atgcaccacg ctacgtagat caggaagctt aactctagcg      600

acctgtcgct cgccccacag aacctcaccc gagaaccaca cattacacgc cgccagctcc      660

cactatactc atcttgcttc ccttaagcgt tctcacgatt cgttcgctgc ccttcttcaa      720

gagtcttctg attctaattc tcattcgaaa tcctctacag ttaatgaatt gcttgacatg      780

acattcattg tctcatggtt ttggcttttt ggcttttgtc ttttaaagct atatcaactt      840

tacatataaa tatacgtcaa aaggggattc attaattaga aaattctctt tttcaatagt      900

tgctattcat tatcaatcta ttcaactcaa ttggttatta ttttcatctt tttgtcatcc      960

taaaccatca acaatattta aatatatctg ttgctacatt aagagttact tcagaaataa     1020

caaaaaaatc gatcaagaat taataaaaat ggactacgct aacatcttga ctgccattcc     1080

tttggaattc accccacaag atgacattgt cttgttggaa ccataccact acttaggtaa     1140

gaacccaggt aaggaaatca gatctcaatt gattgaagct ttcaactact ggttagatgt     1200

caagaaggaa gacttggaag ttatccaaaa tgttgttggt atgttgcaca ccgcttcttt     1260

gttgatggat gatgttgaag attcttccgt cttgagaaga ggttctccag ttgctcattt     1320

gatctacggt attccacaaa ccatcaacac tgctaactac gtttacttct tggcttacca     1380

agaaatcttc aaattgcgtc caactccaat tccaatgcca gttatcccac catcttctgc     1440

ttctttgcaa tcttctgtct cctccgcctc ctcttcctct tctgcctcct ctgaaaacgg     1500

tggtacctcc actccaaact cccaaatccc attctccaag gacacctact tggacaaggt     1560

tatcactgac gaaatgttgt ctttgcaccg tggtcaaggt ttggaattat tctggagaga     1620

ctctttgacc tgtccatctg aagaagaata cgtcaagatg gtcttgggta agaccggtgg     1680

tttgttcaga attgctgtca gattgatgat ggccaagtct gaatgtgaca ttgactttgt     1740

tcaattggtt aacttgattt ccatctactt ccaaatcaga gatgactaca tgaacttgca     1800

atcctctgaa tacgctcaca acaagaactt cgctgaagac ttgactgaag gtaagttctc     1860

cttcccaacc attcactcca ttcacgctaa cccatcttcc agattggtta tcaacacttt     1920

acaaaagaag tccacttctc cagaaatctt acatcactgt gtcaactaca tgagaactga     1980

aacccactct ttcgaataca ctcaagaagt cttgaacact ttatctggtg ctttggaaag     2040

agaattgggt agattacaag gtgaatttgc tgaagctaac tccaagatcg atttgggtga     2100

cgttgaatct gaaggtagaa ccggtaagaa cgtcaaattg gaagccatct tgaagaaatt     2160

ggctgatatc cctctataaa gtgaatttac tttaaatctt gcatttaaat aaattttctt     2220

tttatagctt tatgacttag tttcaattta tatactattt taatgacatt ttcgattcat     2280

tgattgaaag ctttgtgttt tttcttgatg cgctattgca ttgttcttgt ctttttcgcc     2340

acatgtaata tctgtagtag atacctgata cattgtggat gctgagtgaa attttagtta     2400

ataatggagg cgctcttaat aattttgggg atattggctt ttttttttaa agtttacaaa     2460

tgaatttttt ccgccaggat gaagtatgga tctttgaaac ccttttattt tattttactt     2520

tttttagaat                                                            2530


<210>  49
<211>  2130
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT1 - Strong p (ScFBA1p) - crtE - 
       ScTDH3t  -Homology to INT1

<400>  49
catgcctatc cggcggcaac attattactc tggtatacgg gcgtaactcc acttggcttc       60

acatacgttg catacgtcga tatagataat aatgataatg acagcaggat tatcgtaata      120

cgtaatagtt gaaaatctca aaaatgtgtg ggtcattacg taaataatga taggaatggg      180

attcttctat ttttcctttt tccattctag cagccgtcgg gaaaacgtgg catcctctct      240

ttcgggctca attggagtca cgctgccgtg agcatcctct ctttccatat ctaacaactg      300

agcacgtaac caatggaaaa gcatgagctt agcgttgctc caaaaaagta ttggatggtt      360

aataccattt gtctgttctc ttctgacttt gactcctcaa aaaaaaaaaa tctacaatca      420

acagatcgct tcaattacgc cctcacaaaa acttttttcc ttcttcttcg cccacgttaa      480

attttatccc tcatgttgtc taacggattt ctgcacttga tttattataa aaagacaaag      540

acataatact tctctatcaa tttcagttat tgttcttcct tgcgttattc ttctgttctt      600

ctttttcttt tgtcatatat aaccataacc aagtaataca tattcaaaat ggactacgct      660

aacatcttga ctgccattcc tttggaattc accccacaag atgacattgt cttgttggaa      720

ccataccact acttaggtaa gaacccaggt aaggaaatca gatctcaatt gattgaagct      780

ttcaactact ggttagatgt caagaaggaa gacttggaag ttatccaaaa tgttgttggt      840

atgttgcaca ccgcttcttt gttgatggat gatgttgaag attcttccgt cttgagaaga      900

ggttctccag ttgctcattt gatctacggt attccacaaa ccatcaacac tgctaactac      960

gtttacttct tggcttacca agaaatcttc aaattgcgtc caactccaat tccaatgcca     1020

gttatcccac catcttctgc ttctttgcaa tcttctgtct cctccgcctc ctcttcctct     1080

tctgcctcct ctgaaaacgg tggtacctcc actccaaact cccaaatccc attctccaag     1140

gacacctact tggacaaggt tatcactgac gaaatgttgt ctttgcaccg tggtcaaggt     1200

ttggaattat tctggagaga ctctttgacc tgtccatctg aagaagaata cgtcaagatg     1260

gtcttgggta agaccggtgg tttgttcaga attgctgtca gattgatgat ggccaagtct     1320

gaatgtgaca ttgactttgt tcaattggtt aacttgattt ccatctactt ccaaatcaga     1380

gatgactaca tgaacttgca atcctctgaa tacgctcaca acaagaactt cgctgaagac     1440

ttgactgaag gtaagttctc cttcccaacc attcactcca ttcacgctaa cccatcttcc     1500

agattggtta tcaacacttt acaaaagaag tccacttctc cagaaatctt acatcactgt     1560

gtcaactaca tgagaactga aacccactct ttcgaataca ctcaagaagt cttgaacact     1620

ttatctggtg ctttggaaag agaattgggt agattacaag gtgaatttgc tgaagctaac     1680

tccaagatcg atttgggtga cgttgaatct gaaggtagaa ccggtaagaa cgtcaaattg     1740

gaagccatct tgaagaaatt ggctgatatc cctctataaa gtgaatttac tttaaatctt     1800

gcatttaaat aaattttctt tttatagctt tatgacttag tttcaattta tatactattt     1860

taatgacatt ttcgattcat tgattgaaag ctttgtgttt tttcttgatg cgctattgca     1920

ttgttcttgt ctttttcgcc acatgtaata tctgtagtag atacctgata cattgtggat     1980

gctgagtgaa attttagtta ataatggagg cgctcttaat aattttgggg atattggctt     2040

ttttttttaa agtttacaaa tgaatttttt ccgccaggat gaagtatgga tctttgaaac     2100

ccttttattt tattttactt tttttagaat                                      2130


<210>  50
<211>  3428
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT2 - Low p (KlYDR1p) - crtYB - 
       ScPDC1t - Homology to INT2

<400>  50
caaaccgcag attgcgtcac ggttctgcgg acgccgttag cccattggtg tcaagttttc       60

tttttttgcg gtcaccccca tgtggcgggg aggcagagga gtaggtagag caacgaatcc      120

tactatttat ccaaattagt ctaggaactc tttttctaga ttttttagat ttgagggcaa      180

gcgctgttaa cgactcagaa atgtaagcac tacggagtag aacgagaaat ccgccatagg      240

tggaaatcct agcaaaatct tgcttaccct agctagcctc aggtaagcta gccttagcct      300

gtcaaatttt tttcaaaatt tggtaagttt ctactagcaa agcaaacacg gttcaacaaa      360

ccgaaaactc cactcattat acgtggaaac cgaaacaaaa aaacaaaaac caaaatactc      420

gccaatgaga aagttgctgc gtttctactt tcgaggaaga ggaactgaga ggattgacta      480

cgaaaggggc aaaaacgagt cgtattctcc cattattgtc tgctaccacg cggtctagta      540

gaataagcaa ccagtcaacg ctaagacagg taatcaaaat accagtctgc tggctacggg      600

ctagttttta cctcttttag aacccactgt aaaagtccgt tgtaaagccc gttctcactg      660

ttggcgtttt tttttttttg gtttagtttc ttatttttca tttttttctt tcatgaccaa      720

aaacaaacaa atctcgcgat ttgtactgcg gccactgggg cgtggccaaa aaaatgacaa      780

atttagaaac cttagtttct gatttttcct gttatgagga gatatgataa aaaatattac      840

tgctttattg tttttttttt atctactgaa atagagaaac ttacccaagg aggaggcaaa      900

aaaaagagta tatatacagc aggtaccatt cagattttaa tatattcttt tctcttcttc      960

tacactatta ttataataat tttactatat tcatttttag cttaaaacct catagaatat     1020

tattcttcag tcactcgctt aaatacttat caaaaatgac cgctttggct tactaccaaa     1080

tccacttgat ctacactttg ccaatcttag gtttgctagg tttgttgact tctccaattt     1140

tgaccaaatt cgacatctac aagatttcta tcttagtctt tattgctttc tctgctacca     1200

ctccatggga ctcctggatc atcagaaacg gtgcctggac ctacccatct gctgaatctg     1260

gtcaaggtgt tttcggtacc tttttggatg tcccatacga agaatacgcc ttctttgtta     1320

tccaaaccgt catcaccggt ttggtttacg ttttggctac cagacatttg ttgccatctt     1380

tggctctacc aaagacccgt tcttctgcct tgtctctagc tttgaaggct ttaatcccat     1440

tgccaatcat ctatttgttc accgctcatc catctccatc cccagatcct ttggttactg     1500

accactactt ctacatgaga gctttgtctt tgttgatcac cccaccaacc atgttgttgg     1560

ctgctttatc tggtgaatac gctttcgact ggaaatctgg tagagctaag tccaccattg     1620

ctgccatcat gatcccaact gtctacttga tctgggttga ctacgttgcc gttggtcaag     1680

actcctggtc catcaacgat gaaaagattg tcggttggag attaggtggt gtcttgccaa     1740

ttgaagaagc tatgttcttc ttattgacca acttgatgat cgttttgggt ttgtctgcct     1800

gtgaccacac tcaagccttg tacttgttgc acggtagaac tatctacggt aacaagaaga     1860

tgccatcttc tttcccatta atcactccac cagttttgtc cttgttcttc tcctccagac     1920

catactcctc ccaaccaaag agagatttgg aattggctgt caagttgttg gaaaagaagt     1980

ccagatcttt cttcgttgct tctgccggtt tcccatctga agtcagagaa agattggttg     2040

gtttgtacgc tttctgtcgt gtcaccgatg acttgattga ctctccagaa gtttcctcca     2100

acccacacgc taccattgac atggtttccg atttcttgac tttattattc ggtcctccat     2160

tgcacccatc tcaaccagac aagattttgt cttctccatt attaccacct tcccacccat     2220

ccagaccaac tggtatgtac ccattaccac cacctccatc tttgtctcca gctgaattgg     2280

tccaattctt gactgaacgt gtcccagttc aataccactt cgctttcaga ttgttggcca     2340

aattgcaagg tttgattcca agatacccat tggatgaatt attgagaggt tacaccactg     2400

acttgatctt cccattgtcc actgaagccg tccaagctag aaagacccca attgaaacta     2460

ctgctgactt gttggactac ggtttgtgtg ttgccggttc tgttgctgaa ttgttggtct     2520

acgtttcctg ggcttccgct ccatcccaag ttccagctac tattgaagaa agagaagctg     2580

ttttggtcgc ctctcgtgaa atgggtaccg ctttgcaatt ggtcaacatt gccagagata     2640

tcaagggtga cgctactgaa ggtagattct acttgccatt gtctttcttt ggtttgagag     2700

atgaatccaa attggccatt ccaactgact ggactgaacc aagacctcaa gatttcgaca     2760

aattgttgtc tctatctcca tcttccactt taccatcctc taacgcttct gaatccttca     2820

gattcgaatg gaagacctac tctttgccat tggttgctta cgctgaagat ttggctaagc     2880

actcttacaa gggtattgac agattaccaa ctgaagtcca agctggtatg agagctgctt     2940

gtgcttctta cttgttgatt ggtcgtgaaa tcaaggttgt ctggaagggt gatgtcggtg     3000

aaagaagaac cgttgctggt tggagaagag tcagaaaggt tttgtctgtt gtcatgtccg     3060

gttgggaagg tcaataaagc gatttaatct ctaattatta gttaaagttt tataagcatt     3120

tttatgtaac gaaaaataaa ttggttcata ttattactgc actgtcactt accatggaaa     3180

gaccagacaa gaagttgccg acagtctgtt gaattggcct ggttaggctt aagtctgggt     3240

ccgcttcttt acaaatttgg agaatttctc ttaaacgata tgtatattct tttcgttgga     3300

aaagatgtct tccaaaaaaa aaaccgatga attagtggaa ccaaggaaaa aaaaagaggt     3360

atccttgatt aaggaacaga acaatactga ttgcgttacc ccaagcttat cccacgtagt     3420

tttgtgtc                                                              3428


<210>  51
<211>  3420
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT2 - Med p (KlTEF2p) - crtYB - 
       ScPDC1t - Homology to INT2

<400>  51
cgcagattgc gtcacggttc tgcggacgcc gttagcccat tggtgtcaag cctgtccaag       60

caaatgcctt ctcataaatg gtgccaaaga cccgcaagcc caaagcaatt accccccaaa      120

aagaaatgat atagtgcaag atacgtatat gaccatgact tgactaggtg aaacagtgca      180

gaaacagccg cacaaaagca gccctaaccc tcagagtcga ttttactctt tcaggtaata      240

aagcctcgac atcaatttta gacagaagcc aggctggcct cgagattata gccataggca      300

agcaagagga gagaagggga ggccccccat ggggggcctc ccccccgctg tcaaggtttg      360

gcagaaccta gcttcattag gccactagcc cagcctaaaa cgtcaacggg caggaggaac      420

actcccacaa gacggcgtag tattctcgat tcataaccat tttctcaatc gaattacaca      480

gaacacaccg tacaaacctc tctatcataa ctacttaata gtcacacacg tactcgtcta      540

aatacacatc atcgtcctac aagttcatca aagtgttgga cagacaacta taccagcatg      600

gatctcttgt atcggttctt ttctcccgct ctctcgcaat aacaatgaac actgggtcaa      660

tcatagccta cacaggtgaa cagagtagcg tttatacagg gtttatacgg tgattcctac      720

ggcaaaaatt tttcatttct aaaaaaaaaa agaaaaattt ttctttccaa cgctagaagg      780

aaaagaaaaa tctaattaaa ttgatttggt gattttctga gagttccctt tttcatatat      840

cgaattttga atataaaagg agatcgaaaa aatttttcta ttcaatctgt tttctggttt      900

tatttgatag tttttttgtg tattattatt atggattagt actggtttat atgggttttt      960

ctgtataact tctttttatt ttagtttgtt taatcttatt ttgagttaca ttatagttcc     1020

ctaactgcaa gagaagtaac attaaaaatg accgctttgg cttactacca aatccacttg     1080

atctacactt tgccaatctt aggtttgcta ggtttgttga cttctccaat tttgaccaaa     1140

ttcgacatct acaagatttc tatcttagtc tttattgctt tctctgctac cactccatgg     1200

gactcctgga tcatcagaaa cggtgcctgg acctacccat ctgctgaatc tggtcaaggt     1260

gttttcggta cctttttgga tgtcccatac gaagaatacg ccttctttgt tatccaaacc     1320

gtcatcaccg gtttggttta cgttttggct accagacatt tgttgccatc tttggctcta     1380

ccaaagaccc gttcttctgc cttgtctcta gctttgaagg ctttaatccc attgccaatc     1440

atctatttgt tcaccgctca tccatctcca tccccagatc ctttggttac tgaccactac     1500

ttctacatga gagctttgtc tttgttgatc accccaccaa ccatgttgtt ggctgcttta     1560

tctggtgaat acgctttcga ctggaaatct ggtagagcta agtccaccat tgctgccatc     1620

atgatcccaa ctgtctactt gatctgggtt gactacgttg ccgttggtca agactcctgg     1680

tccatcaacg atgaaaagat tgtcggttgg agattaggtg gtgtcttgcc aattgaagaa     1740

gctatgttct tcttattgac caacttgatg atcgttttgg gtttgtctgc ctgtgaccac     1800

actcaagcct tgtacttgtt gcacggtaga actatctacg gtaacaagaa gatgccatct     1860

tctttcccat taatcactcc accagttttg tccttgttct tctcctccag accatactcc     1920

tcccaaccaa agagagattt ggaattggct gtcaagttgt tggaaaagaa gtccagatct     1980

ttcttcgttg cttctgccgg tttcccatct gaagtcagag aaagattggt tggtttgtac     2040

gctttctgtc gtgtcaccga tgacttgatt gactctccag aagtttcctc caacccacac     2100

gctaccattg acatggtttc cgatttcttg actttattat tcggtcctcc attgcaccca     2160

tctcaaccag acaagatttt gtcttctcca ttattaccac cttcccaccc atccagacca     2220

actggtatgt acccattacc accacctcca tctttgtctc cagctgaatt ggtccaattc     2280

ttgactgaac gtgtcccagt tcaataccac ttcgctttca gattgttggc caaattgcaa     2340

ggtttgattc caagataccc attggatgaa ttattgagag gttacaccac tgacttgatc     2400

ttcccattgt ccactgaagc cgtccaagct agaaagaccc caattgaaac tactgctgac     2460

ttgttggact acggtttgtg tgttgccggt tctgttgctg aattgttggt ctacgtttcc     2520

tgggcttccg ctccatccca agttccagct actattgaag aaagagaagc tgttttggtc     2580

gcctctcgtg aaatgggtac cgctttgcaa ttggtcaaca ttgccagaga tatcaagggt     2640

gacgctactg aaggtagatt ctacttgcca ttgtctttct ttggtttgag agatgaatcc     2700

aaattggcca ttccaactga ctggactgaa ccaagacctc aagatttcga caaattgttg     2760

tctctatctc catcttccac tttaccatcc tctaacgctt ctgaatcctt cagattcgaa     2820

tggaagacct actctttgcc attggttgct tacgctgaag atttggctaa gcactcttac     2880

aagggtattg acagattacc aactgaagtc caagctggta tgagagctgc ttgtgcttct     2940

tacttgttga ttggtcgtga aatcaaggtt gtctggaagg gtgatgtcgg tgaaagaaga     3000

accgttgctg gttggagaag agtcagaaag gttttgtctg ttgtcatgtc cggttgggaa     3060

ggtcaataaa gcgatttaat ctctaattat tagttaaagt tttataagca tttttatgta     3120

acgaaaaata aattggttca tattattact gcactgtcac ttaccatgga aagaccagac     3180

aagaagttgc cgacagtctg ttgaattggc ctggttaggc ttaagtctgg gtccgcttct     3240

ttacaaattt ggagaatttc tcttaaacga tatgtatatt cttttcgttg gaaaagatgt     3300

cttccaaaaa aaaaaccgat gaattagtgg aaccaaggaa aaaaaaagag gtatccttga     3360

ttaaggaaca gaacaatact gattgcgtta ccccaagctt atcccacgta gttttgtgtc     3420


<210>  52
<211>  3025
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT2 - Strong p (ScTEF1p) - crtYB - 
       ScPDC1t - Homology to INT2

<400>  52
caaaccgcag attgcgtcac ggttctgcgg acgccgttag cccattggtg tcaaggctga       60

taatagcgta taaacaatgc atactttgta cgttcaaaat acaatgcagt agatatattt      120

atgcatatta catataatac atatcacata ggaagcaaca ggcgcgttgg acttttaatt      180

ttcgaggacc gcgaatcctt acatcacacc caatccccca caagtgatcc cccacacacc      240

atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca      300

tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc      360

tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaca ccgcctcgtt      420

tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat      480

tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg      540

gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt      600

cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca aaatgaccgc      660

tttggcttac taccaaatcc acttgatcta cactttgcca atcttaggtt tgctaggttt      720

gttgacttct ccaattttga ccaaattcga catctacaag atttctatct tagtctttat      780

tgctttctct gctaccactc catgggactc ctggatcatc agaaacggtg cctggaccta      840

cccatctgct gaatctggtc aaggtgtttt cggtaccttt ttggatgtcc catacgaaga      900

atacgccttc tttgttatcc aaaccgtcat caccggtttg gtttacgttt tggctaccag      960

acatttgttg ccatctttgg ctctaccaaa gacccgttct tctgccttgt ctctagcttt     1020

gaaggcttta atcccattgc caatcatcta tttgttcacc gctcatccat ctccatcccc     1080

agatcctttg gttactgacc actacttcta catgagagct ttgtctttgt tgatcacccc     1140

accaaccatg ttgttggctg ctttatctgg tgaatacgct ttcgactgga aatctggtag     1200

agctaagtcc accattgctg ccatcatgat cccaactgtc tacttgatct gggttgacta     1260

cgttgccgtt ggtcaagact cctggtccat caacgatgaa aagattgtcg gttggagatt     1320

aggtggtgtc ttgccaattg aagaagctat gttcttctta ttgaccaact tgatgatcgt     1380

tttgggtttg tctgcctgtg accacactca agccttgtac ttgttgcacg gtagaactat     1440

ctacggtaac aagaagatgc catcttcttt cccattaatc actccaccag ttttgtcctt     1500

gttcttctcc tccagaccat actcctccca accaaagaga gatttggaat tggctgtcaa     1560

gttgttggaa aagaagtcca gatctttctt cgttgcttct gccggtttcc catctgaagt     1620

cagagaaaga ttggttggtt tgtacgcttt ctgtcgtgtc accgatgact tgattgactc     1680

tccagaagtt tcctccaacc cacacgctac cattgacatg gtttccgatt tcttgacttt     1740

attattcggt cctccattgc acccatctca accagacaag attttgtctt ctccattatt     1800

accaccttcc cacccatcca gaccaactgg tatgtaccca ttaccaccac ctccatcttt     1860

gtctccagct gaattggtcc aattcttgac tgaacgtgtc ccagttcaat accacttcgc     1920

tttcagattg ttggccaaat tgcaaggttt gattccaaga tacccattgg atgaattatt     1980

gagaggttac accactgact tgatcttccc attgtccact gaagccgtcc aagctagaaa     2040

gaccccaatt gaaactactg ctgacttgtt ggactacggt ttgtgtgttg ccggttctgt     2100

tgctgaattg ttggtctacg tttcctgggc ttccgctcca tcccaagttc cagctactat     2160

tgaagaaaga gaagctgttt tggtcgcctc tcgtgaaatg ggtaccgctt tgcaattggt     2220

caacattgcc agagatatca agggtgacgc tactgaaggt agattctact tgccattgtc     2280

tttctttggt ttgagagatg aatccaaatt ggccattcca actgactgga ctgaaccaag     2340

acctcaagat ttcgacaaat tgttgtctct atctccatct tccactttac catcctctaa     2400

cgcttctgaa tccttcagat tcgaatggaa gacctactct ttgccattgg ttgcttacgc     2460

tgaagatttg gctaagcact cttacaaggg tattgacaga ttaccaactg aagtccaagc     2520

tggtatgaga gctgcttgtg cttcttactt gttgattggt cgtgaaatca aggttgtctg     2580

gaagggtgat gtcggtgaaa gaagaaccgt tgctggttgg agaagagtca gaaaggtttt     2640

gtctgttgtc atgtccggtt gggaaggtca ataaagcgat ttaatctcta attattagtt     2700

aaagttttat aagcattttt atgtaacgaa aaataaattg gttcatatta ttactgcact     2760

gtcacttacc atggaaagac cagacaagaa gttgccgaca gtctgttgaa ttggcctggt     2820

taggcttaag tctgggtccg cttctttaca aatttggaga atttctctta aacgatatgt     2880

atattctttt cgttggaaaa gatgtcttcc aaaaaaaaaa ccgatgaatt agtggaacca     2940

aggaaaaaaa aagaggtatc cttgattaag gaacagaaca atactgattg cgttacccca     3000

agcttatccc acgtagtttt gtgtc                                           3025


<210>  53
<211>  2748
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT3 - Low p (ScPRE3p) - crtI - 
       ScTAL1t - Homology to INT3

<400>  53
gtccttgtat ggaagtatca aaggggacgt tcttcacctc cttggaacaa acattaattt       60

gttctgcata ctttgaacct ttcagaaaat aaaaaacatt acgcgcatac ttaccctgct      120

cgcgaagaag agtaacacta acgcattcta tgggcaattg aagacagtat tcagtacaag      180

acatagtccg tttccttgag tcaattccta tagcattatg aactagccgc ctttaagagt      240

gccaagctgt tcaacaccga tcatttttga tgatttggcg tttttgttat attgatagat      300

ttcttttgaa ttttgtcatt ttcacttttc cactcgcaac ggaatccggt ggcaaaaaag      360

ggaaaagcat tgaaatgcaa tctttaacag tattttaaac aagttgcgac acggtgtaca      420

attacgataa gaattgctac ttcaaagtac acacagaaag ttaacatgaa tggaattcaa      480

gtggacatca atcgtttgaa aaagggcgaa gtcagtttag gtacctcaat gtatgtatat      540

aagaattttt cctcccactt tattgtttct aaaagttcaa tgaagtaaag tctcaattgg      600

ccttattact aactaatagg tatcttataa tcacctaata aaatagaatg ggtaaggaac      660

aagaccaaga caagccaact gccatcatcg ttggttgtgg tatcggtggt attgctaccg      720

ctgccagatt agctaaggaa ggtttccaag ttaccgtctt tgaaaagaac gactactccg      780

gtggtagatg ttctttgatt gaaagagatg gttacagatt cgaccaaggt ccatctttgt      840

tgctattacc agacttgttc aagcaaacct tcgaagattt gggtgaaaag atggaagact      900

gggttgattt gatcaagtgt gaaccaaact acgtttgtca cttccatgat gaagaaactt      960

tcaccttctc cactgacatg gctttattga agagagaagt cgaaagattt gaaggtaaag     1020

atggtttcga cagattcttg tctttcatcc aagaagctca cagacattac gaattggctg     1080

ttgtccacgt cttgcaaaag aacttcccag gtttcgctgc tttcttgaga ttacaattca     1140

tcggtcaaat cttagctttg cacccatttg aatccatctg gaccagagtt tgtcgttact     1200

tcaagactga cagattgaga agagtcttct cctttgccgt tatgtacatg ggtcaatctc     1260

catactctgc tccaggtacc tactccttgt tgcaatacac tgaattgact gaaggtatct     1320

ggtacccaag aggtggtttc tggcaagttc caaacacttt gttgcaaatc gtcaagagaa     1380

acaacccatc tgctaagttc aacttcaacg ctccagtttc tcaagttttg ttgtctccag     1440

ctaaggacag agctaccggt gtcagattag aatctggtga agaacaccac gctgatgttg     1500

tcattgtcaa tgctgacttg gtctacgctt ctgaacattt gattccagat gatgctagaa     1560

acaagatcgg tcaattaggt gaagttaagc gttcctggtg ggctgatttg gttggtggta     1620

agaagttgaa gggttcttgt tcttctttgt ctttctactg gtctatggac agaatcgttg     1680

acggtttggg tggtcacaac atcttcttgg ctgaagactt caagggttcc ttcgacacca     1740

ttttcgaaga attgggtttg ccagctgacc catctttcta tgttaacgtt ccatccagaa     1800

ttgacccttc tgctgctcca gaaggtaagg atgccattgt catcttagtc ccatgtggtc     1860

acatcgatgc ttccaaccct caagactaca acaaattggt tgccagagcc agaaagttcg     1920

tcatccaaac cttgtctgcc aagttgggtc taccagattt cgaaaagatg attgttgctg     1980

aaaaggttca cgatgctcca tcctgggaaa aggaattcaa cttgaaggac ggttccattt     2040

tgggtttggc tcacaacttc atgcaagtct tgggtttcag accatccacc agacacccaa     2100

agtacgacaa attgttcttt gtcggtgctt ctacccaccc aggtactggt gttccaattg     2160

tcttggctgg tgccaaattg actgctaacc aagttttgga atccttcgat cgttctccag     2220

ctccagatcc taacatgtct ttgtctgttc catacggtaa gccattgaaa tccaacggta     2280

ctggtattga ctctcaagtc caattgaaat tcatggactt ggaacgttgg gtttacctat     2340

tagtcttgtt gattggtgct gttatcgcca gatccgtcgg tgtcttggcc ttttaaagga     2400

agtatctcgg aaatattaat ttaggccatg tccttatgca cgtttctttt gatacttacg     2460

ggtacatgta cacaagtata tctatatata taaattaatg aaaatcccct atttatatat     2520

atgactttaa cgagacagaa cagtttttta ttttttatcc tatttgatga atgatacagt     2580

ttcttattca cgtgttatac ccacaccaaa tccaatagca ataccggcca tcacaatcac     2640

tgtttcggca gcccctaaga tcagacaaaa catccggaac caccttaaat caacgtcgat     2700

gggacgtcag cactgtactt gtttttgcga ctagattgta aatcattc                  2748


<210>  54
<211>  2749
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT3 - Med p (ScACT1p) - crtI - 
       ScTAL1t - Homology to INT3

<400>  54
cgtccttgta tggaagtatc aaaggggacg ttcttcacct ccttggaaaa catatataca       60

caattacagt aacaataaca agaggacaga tactaccaaa atgtgtgggg aagcgggtaa      120

gctgccacag caattaatgc acaacattta acctacattc ttccttatcg gatcctcaaa      180

acccttaaaa acatatgcct caccctaaca tattttccaa ttaaccctca atatttctct      240

gtcacccggc ctctattttc cattttcttc tttacccgcc acgcgttttt ttctttcaaa      300

tttttttctt ctttcttctt tttcttccac gtcctcttgc ataaataaat aaaccgtttt      360

gaaaccaaac tcgcctctct ctctcctttt tgaaatattt ttgggtttgt ttgatccttt      420

ccttcccaat ctctcttgtt taatatatat tcatttatat cacgctctct ttttatcttc      480

ctttttttcc tctctcttgt attcttcctt cccctttcta ctcaaaccaa gaagaaaaag      540

aaaaggtcaa tctttgttaa agaataggat cttctactac atcagctttt agatttttca      600

cgcttactgc ttttttcttc ccaagatcga aaatttactg aattaacaat gggtaaggaa      660

caagaccaag acaagccaac tgccatcatc gttggttgtg gtatcggtgg tattgctacc      720

gctgccagat tagctaagga aggtttccaa gttaccgtct ttgaaaagaa cgactactcc      780

ggtggtagat gttctttgat tgaaagagat ggttacagat tcgaccaagg tccatctttg      840

ttgctattac cagacttgtt caagcaaacc ttcgaagatt tgggtgaaaa gatggaagac      900

tgggttgatt tgatcaagtg tgaaccaaac tacgtttgtc acttccatga tgaagaaact      960

ttcaccttct ccactgacat ggctttattg aagagagaag tcgaaagatt tgaaggtaaa     1020

gatggtttcg acagattctt gtctttcatc caagaagctc acagacatta cgaattggct     1080

gttgtccacg tcttgcaaaa gaacttccca ggtttcgctg ctttcttgag attacaattc     1140

atcggtcaaa tcttagcttt gcacccattt gaatccatct ggaccagagt ttgtcgttac     1200

ttcaagactg acagattgag aagagtcttc tcctttgccg ttatgtacat gggtcaatct     1260

ccatactctg ctccaggtac ctactccttg ttgcaataca ctgaattgac tgaaggtatc     1320

tggtacccaa gaggtggttt ctggcaagtt ccaaacactt tgttgcaaat cgtcaagaga     1380

aacaacccat ctgctaagtt caacttcaac gctccagttt ctcaagtttt gttgtctcca     1440

gctaaggaca gagctaccgg tgtcagatta gaatctggtg aagaacacca cgctgatgtt     1500

gtcattgtca atgctgactt ggtctacgct tctgaacatt tgattccaga tgatgctaga     1560

aacaagatcg gtcaattagg tgaagttaag cgttcctggt gggctgattt ggttggtggt     1620

aagaagttga agggttcttg ttcttctttg tctttctact ggtctatgga cagaatcgtt     1680

gacggtttgg gtggtcacaa catcttcttg gctgaagact tcaagggttc cttcgacacc     1740

attttcgaag aattgggttt gccagctgac ccatctttct atgttaacgt tccatccaga     1800

attgaccctt ctgctgctcc agaaggtaag gatgccattg tcatcttagt cccatgtggt     1860

cacatcgatg cttccaaccc tcaagactac aacaaattgg ttgccagagc cagaaagttc     1920

gtcatccaaa ccttgtctgc caagttgggt ctaccagatt tcgaaaagat gattgttgct     1980

gaaaaggttc acgatgctcc atcctgggaa aaggaattca acttgaagga cggttccatt     2040

ttgggtttgg ctcacaactt catgcaagtc ttgggtttca gaccatccac cagacaccca     2100

aagtacgaca aattgttctt tgtcggtgct tctacccacc caggtactgg tgttccaatt     2160

gtcttggctg gtgccaaatt gactgctaac caagttttgg aatccttcga tcgttctcca     2220

gctccagatc ctaacatgtc tttgtctgtt ccatacggta agccattgaa atccaacggt     2280

actggtattg actctcaagt ccaattgaaa ttcatggact tggaacgttg ggtttaccta     2340

ttagtcttgt tgattggtgc tgttatcgcc agatccgtcg gtgtcttggc cttttaaagg     2400

aagtatctcg gaaatattaa tttaggccat gtccttatgc acgtttcttt tgatacttac     2460

gggtacatgt acacaagtat atctatatat ataaattaat gaaaatcccc tatttatata     2520

tatgacttta acgagacaga acagtttttt attttttatc ctatttgatg aatgatacag     2580

tttcttattc acgtgttata cccacaccaa atccaatagc aataccggcc atcacaatca     2640

ctgtttcggc agcccctaag atcagacaaa acatccggaa ccaccttaaa tcaacgtcga     2700

tgggacgtca gcactgtact tgtttttgcg actagattgt aaatcattc                 2749


<210>  55
<211>  3149
<212>  DNA
<213>  artificial sequence

<220>
<223>  donor expression cassette sequence with 50 bp LF and RF flanks 
       (PCR fragment). Homology to INT3 - Strong p (KlENO1p) - crtI - 
       ScTAL1t - Homology to INT3

<400>  55
cgtccttgta tggaagtatc aaaggggacg ttcttcacct ccttggaacg tatccctatc       60

tggattaaca tcactgccac agatcgaatt gcaagaagcc acacttcacg tgatccactc      120

gttcatcagg tttgtagctt catggcgcag gacttctgat gatgaactat ctggctcatc      180

cggatggatc acaaggatgg ataccctcag acagtacgtt tccgtatgga gcgatcttac      240

aagaaccaac agttgatcct attacttttt ttttattttt tgtccctccg ggatggcaag      300

agggacaaag aagaatcttc gttcttcttt cttgttctca acttcccagc ttccgtgtga      360

ttaccctccg ggacaacaga aaaactggca ttcggtatcc cgggaatctg ctgagaagga      420

aagaaaacga aaaaaaaatt gtacatttgt gtcacattat gaattacagg aagtcagaaa      480

acaggcagca catgtctcgc acatgcatgt ccatcagacg agacattatg agacatgcac      540

gcgtgtgaga gacatagcaa aagtctctcc agtacacaca gaaagacacg ttcacaatcc      600

aggcacccca cagagaaaaa aaaaagaaga agcccggaag ctggcacgcc atcatcaacc      660

accgctcggt ttacacgcat cccaactgtc ttttttttct ggaatcctat aataactggc      720

atctggaaat cacgttgtat gttgcaccat agtgactggc tgtctgacta gcaaacattg      780

attccctgat tcccatttgg ctcaattttg atgagaaaca gttgattgat tcttgtcaat      840

ttttttttct ttggaccacc accaaccaat tgacattgaa gtactttccc atgatttgag      900

gttatataaa aggacgttca aatcactttc aaggttaatt cagttttgtc aattgattta      960

agttcaattg ttaacaaatt taatttaatt cgaaacaaac caaaccaatt catttgaatt     1020

aacaaaccaa cccacaaaac aaaaaaaaat gggtaaggaa caagaccaag acaagccaac     1080

tgccatcatc gttggttgtg gtatcggtgg tattgctacc gctgccagat tagctaagga     1140

aggtttccaa gttaccgtct ttgaaaagaa cgactactcc ggtggtagat gttctttgat     1200

tgaaagagat ggttacagat tcgaccaagg tccatctttg ttgctattac cagacttgtt     1260

caagcaaacc ttcgaagatt tgggtgaaaa gatggaagac tgggttgatt tgatcaagtg     1320

tgaaccaaac tacgtttgtc acttccatga tgaagaaact ttcaccttct ccactgacat     1380

ggctttattg aagagagaag tcgaaagatt tgaaggtaaa gatggtttcg acagattctt     1440

gtctttcatc caagaagctc acagacatta cgaattggct gttgtccacg tcttgcaaaa     1500

gaacttccca ggtttcgctg ctttcttgag attacaattc atcggtcaaa tcttagcttt     1560

gcacccattt gaatccatct ggaccagagt ttgtcgttac ttcaagactg acagattgag     1620

aagagtcttc tcctttgccg ttatgtacat gggtcaatct ccatactctg ctccaggtac     1680

ctactccttg ttgcaataca ctgaattgac tgaaggtatc tggtacccaa gaggtggttt     1740

ctggcaagtt ccaaacactt tgttgcaaat cgtcaagaga aacaacccat ctgctaagtt     1800

caacttcaac gctccagttt ctcaagtttt gttgtctcca gctaaggaca gagctaccgg     1860

tgtcagatta gaatctggtg aagaacacca cgctgatgtt gtcattgtca atgctgactt     1920

ggtctacgct tctgaacatt tgattccaga tgatgctaga aacaagatcg gtcaattagg     1980

tgaagttaag cgttcctggt gggctgattt ggttggtggt aagaagttga agggttcttg     2040

ttcttctttg tctttctact ggtctatgga cagaatcgtt gacggtttgg gtggtcacaa     2100

catcttcttg gctgaagact tcaagggttc cttcgacacc attttcgaag aattgggttt     2160

gccagctgac ccatctttct atgttaacgt tccatccaga attgaccctt ctgctgctcc     2220

agaaggtaag gatgccattg tcatcttagt cccatgtggt cacatcgatg cttccaaccc     2280

tcaagactac aacaaattgg ttgccagagc cagaaagttc gtcatccaaa ccttgtctgc     2340

caagttgggt ctaccagatt tcgaaaagat gattgttgct gaaaaggttc acgatgctcc     2400

atcctgggaa aaggaattca acttgaagga cggttccatt ttgggtttgg ctcacaactt     2460

catgcaagtc ttgggtttca gaccatccac cagacaccca aagtacgaca aattgttctt     2520

tgtcggtgct tctacccacc caggtactgg tgttccaatt gtcttggctg gtgccaaatt     2580

gactgctaac caagttttgg aatccttcga tcgttctcca gctccagatc ctaacatgtc     2640

tttgtctgtt ccatacggta agccattgaa atccaacggt actggtattg actctcaagt     2700

ccaattgaaa ttcatggact tggaacgttg ggtttaccta ttagtcttgt tgattggtgc     2760

tgttatcgcc agatccgtcg gtgtcttggc cttttaaagg aagtatctcg gaaatattaa     2820

tttaggccat gtccttatgc acgtttcttt tgatacttac gggtacatgt acacaagtat     2880

atctatatat ataaattaat gaaaatcccc tatttatata tatgacttta acgagacaga     2940

acagtttttt attttttatc ctatttgatg aatgatacag tttcttattc acgtgttata     3000

cccacaccaa atccaatagc aataccggcc atcacaatca ctgtttcggc agcccctaag     3060

atcagacaaa acatccggaa ccaccttaaa tcaacgtcga tgggacgtca gcactgtact     3120

tgtttttgcg actagattgt aaatcattc                                       3149


<210>  56
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 47

<400>  56
catgcctatc cggcggcaac attattactc tggtatacgg gcgtaactcc acgtaaaaac       60

taaaacgagc c                                                            71


<210>  57
<211>  74
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 47, 48, 49

<400>  57
attctaaaaa aagtaaaata aaataaaagg gtttcaaaga tccatacttc atcctggcgg       60

aaaaaattca tttg                                                         74


<210>  58
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 48

<400>  58
catgcctatc cggcggcaac attattactc tggtatacgg gcgtaactcc acctcatcac       60

tagaagccga ac                                                           72


<210>  59
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 49

<400>  59
catgcctatc cggcggcaac attattactc tggtatacgg gcgtaactcc acttggcttc       60

acatacgttg c                                                            71


<210>  60
<211>  84
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 50

<400>  60
caaaccgcag attgcgtcac ggttctgcgg acgccgttag cccattggtg tcaagttttc       60

tttttttgcg gtcaccccca tgtg                                              84


<210>  61
<211>  70
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 50, 51, 52

<400>  61
gacacaaaac tacgtgggat aagcttgggg taacgcaatc agtattgttc tgttccttaa       60

tcaaggatac                                                              70


<210>  62
<211>  73
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 51

<400>  62
cgcagattgc gtcacggttc tgcggacgcc gttagcccat tggtgtcaag cctgtccaag       60

caaatgcctt ctc                                                          73


<210>  63
<211>  80
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 52

<400>  63
caaaccgcag attgcgtcac ggttctgcgg acgccgttag cccattggtg tcaaggctga       60

taatagcgta taaacaatgc                                                   80


<210>  64
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 53

<400>  64
gtccttgtat ggaagtatca aaggggacgt tcttcacctc cttggaacaa acattaattt       60

gttctgcata c                                                            71


<210>  65
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 53, 54, 55

<400>  65
gaatgattta caatctagtc gcaaaaacaa gtacagtgct gacgtcccat cgacgttgat       60

ttaaggtggt tc                                                           72


<210>  66
<211>  73
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 54

<400>  66
cgtccttgta tggaagtatc aaaggggacg ttcttcacct ccttggaaaa catatataca       60

caattacagt aac                                                          73


<210>  67
<211>  74
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 55

<400>  67
cgtccttgta tggaagtatc aaaggggacg ttcttcacct ccttggaacg tatccctatc       60

tggattaaca tcac                                                         74


<210>  68
<211>  584
<212>  DNA
<213>  artificial sequence

<220>
<223>  right flank sequence with overlap ScTDH3t - INT1 3' (part of 
       donor) - INT1 3' genomic DNA. Homology with INT1 3' genomic DNA 
       524 bp

<400>  68
aattttgggg atattggctt ttttttttaa agtttacaaa tgaatttttt ccgccaggat       60

gaagtatgga tctttgaaac ccttttattt tattttactt tttttagaat gacctgttcc      120

cgacactatg taagatctag cttttaacat attatggaaa cctgaaatgt aaaatctgaa      180

tttttgtata tgtgtttata tttgggtagt tcttttgagg aaagcatgca tagacttgct      240

gtacgaactt tatgtgactt gtagtgacgc tgtttcatga gactttagcc ctttgaacat      300

attatcatat ctcagcttga aatactatag atttactttt gcagccattt cttggtgctc      360

caaggttgtg cgtatctatt acttaatttc tgtccttgcc aagttttgca gcagggcggt      420

cacaagactc ctctgccgtc attccttagt ccttcgggaa cacacttatt tatgtatttg      480

tattctacaa ttctacggtg cacaagggtt gggcactgtt gagctcagca cgcaactatt      540

gctggcatga agataagatt gatttttgga agaataagct tgtg                       584


<210>  69
<211>  372
<212>  DNA
<213>  artificial sequence

<220>
<223>  left flank sequence with overlap KlYDR2p - INT2 5' (part of 
       donor) - INT2 5' genomic DNA. Homology with INT2 5' genomic DNA 
       322 bp

<400>  69
cattatatcg aggaaagccc tgcgaacgtg aagtgaaaag gacggttgtc ctttttcctg       60

ttcttcgagg atcctgaaca ggatgtcagt agggcgggcc ttttcttttt attgtgcagg      120

tacgcatgtt tctttgacac aaaatagtct ggattcgcca gccagtggca cttttgatgc      180

catttccacc acagaaaacg tctgagtatt cagccatggg aggggagtgt tgaaagggga      240

gaagcagccc acttctctgt cttgacacaa accgcagatt gcgtcacggt tctgcggacg      300

ccgttagccc attggtgtca agttttcttt ttttgcggtc acccccatgt ggcggggagg      360

cagaggagta gg                                                          372


<210>  70
<211>  372
<212>  DNA
<213>  artificial sequence

<220>
<223>  left flank sequence with overlap KlTEF2p - INT2 5' (part of 
       donor) - INT2 5' genomic DNA. Homology with INT2 5' genomic DNA 
       322 bp

<400>  70
cattatatcg aggaaagccc tgcgaacgtg aagtgaaaag gacggttgtc ctttttcctg       60

ttcttcgagg atcctgaaca ggatgtcagt agggcgggcc ttttcttttt attgtgcagg      120

tacgcatgtt tctttgacac aaaatagtct ggattcgcca gccagtggca cttttgatgc      180

catttccacc acagaaaacg tctgagtatt cagccatggg aggggagtgt tgaaagggga      240

gaagcagccc acttctctgt cttgacacaa accgcagatt gcgtcacggt tctgcggacg      300

ccgttagccc attggtgtca agcctgtcca agcaaatgcc ttctcataaa tggtgccaaa      360

gacccgcaag cc                                                          372


<210>  71
<211>  373
<212>  DNA
<213>  artificial sequence

<220>
<223>  left flank sequence with overlap ScTEF1p - INT2 5' (part of 
       donor) - INT2 5' genomic DNA. Homology with INT2 5' genomic DNA 
       322 bp

<400>  71
cattatatcg aggaaagccc tgcgaacgtg aagtgaaaag gacggttgtc ctttttcctg       60

ttcttcgagg atcctgaaca ggatgtcagt agggcgggcc ttttcttttt attgtgcagg      120

tacgcatgtt tctttgacac aaaatagtct ggattcgcca gccagtggca cttttgatgc      180

catttccacc acagaaaacg tctgagtatt cagccatggg aggggagtgt tgaaagggga      240

gaagcagccc acttctctgt cttgacacaa accgcagatt gcgtcacggt tctgcggacg      300

ccgttagccc attggtgtca aggctgataa tagcgtataa acaatgcata ctttgtacgt      360

tcaaaataca atg                                                         373


<210>  72
<211>  608
<212>  DNA
<213>  artificial sequence

<220>
<223>  right flank sequence with overlap ScPDC1t - INT2 3' (part of 
       donor) - INT2 3' genomic DNA. Homology with INT2 3' genomic DNA 
       524 bp

<400>  72
gaattagtgg aaccaaggaa aaaaaaagag gtatccttga ttaaggaaca gaacaatact       60

gattgcgtta ccccaagctt atcccacgta gttttgtgtc tctttcttgg gctcttggcg      120

gccgtacgca tatagtatac taatgcggta acggctacct cctacgtctg tatcaaaaaa      180

gttcgctctg gtcctatcaa tattttctcc cacacgagga ttctgtggtt ttgcagccac      240

atctacagga tgttgacttt ttgtttgttt acgtagtttg tttcaacgct ttcgaagatg      300

tggcgctggt gcaagtcatc taagttggtt actgataaca aattgtgaca aggatgtagt      360

ggatgcgtag gagaccaaaa tgacatctga cacaaaaaaa agtagataac gaaaacagga      420

caaaggtgtc attttgtttt gccaaggaga attataatct tagtgtgaag atactttcat      480

tagacacata aatttacgtt cgacacactt ttttcttact cctcccctta tcaaggatac      540

ctggttgatt ctacgtcgtc ctcttcactt tggttaattc acctttgccc ttcacctcgt      600

ggtgtgcg                                                               608


<210>  73
<211>  649
<212>  DNA
<213>  artificial sequence

<220>
<223>  left flank sequence with overlap ScPRE3p - INT3 5' (part of 
       donor) - INT3 5' genomic DNA. Homology with INT3 5' genomic DNA 
       602 bp

<400>  73
taaaggaggt gcacgcatta tggagaccac tacgatacga tagctgcgtt gttgttgaag       60

gggtttctta aggttgtttt cgttgaaggt aaatattggt cgtttttgtg cagcatattg      120

tcctctagat gcaaactctg caggtccatt tgcagtaaag tgagttgcct ctcgaagaat      180

cattaatttc gtataaccgt cactattaaa gtcagaaaat aaattctgtc gtagacaatg      240

ttaccataat gttcttgtcc attttgcata cactttaaat attcatttga tttctcaggg      300

ttcatgatca taataaattg cgcattcgca aggcggtagt attataatgg ggtccatcat      360

tctgtagcaa gaagttacag tacgctgttc aagcgttaaa caagataagt aatctcgaat      420

gaaacattca tatttcgcat gagccaacat acagttgctg agtaatcttc attgcgctta      480

tttatcggca ttgagattgt aaaggaagta aaacgcattt ttgcagatct gttctcttat      540

gtatttttaa tcgtccttgt atggaagtat caaaggggac gttcttcacc tccttggaac      600

aaacattaat ttgttctgca tactttgaac ctttcagaaa ataaaaaac                  649


<210>  74
<211>  648
<212>  DNA
<213>  artificial sequence

<220>
<223>  left flank sequence with overlap ScACT1p - INT3 5' (part of 
       donor) - INT3 5' genomic DNA. Homology with INT3 5' genomic DNA 
       602 bp

<400>  74
taaaggaggt gcacgcatta tggagaccac tacgatacga tagctgcgtt gttgttgaag       60

gggtttctta aggttgtttt cgttgaaggt aaatattggt cgtttttgtg cagcatattg      120

tcctctagat gcaaactctg caggtccatt tgcagtaaag tgagttgcct ctcgaagaat      180

cattaatttc gtataaccgt cactattaaa gtcagaaaat aaattctgtc gtagacaatg      240

ttaccataat gttcttgtcc attttgcata cactttaaat attcatttga tttctcaggg      300

ttcatgatca taataaattg cgcattcgca aggcggtagt attataatgg ggtccatcat      360

tctgtagcaa gaagttacag tacgctgttc aagcgttaaa caagataagt aatctcgaat      420

gaaacattca tatttcgcat gagccaacat acagttgctg agtaatcttc attgcgctta      480

tttatcggca ttgagattgt aaaggaagta aaacgcattt ttgcagatct gttctcttat      540

gtatttttaa tcgtccttgt atggaagtat caaaggggac gttcttcacc tccttggaaa      600

acatatatac acaattacag taacaataac aagaggacag atactacc                   648


<210>  75
<211>  649
<212>  DNA
<213>  artificial sequence

<220>
<223>  left flank sequence with overlap KlENO1p - INT3 5' (part of 
       donor) - INT3 5' genomic DNA. Homology with INT3 5' genomic DNA 
       602 bp

<400>  75
taaaggaggt gcacgcatta tggagaccac tacgatacga tagctgcgtt gttgttgaag       60

gggtttctta aggttgtttt cgttgaaggt aaatattggt cgtttttgtg cagcatattg      120

tcctctagat gcaaactctg caggtccatt tgcagtaaag tgagttgcct ctcgaagaat      180

cattaatttc gtataaccgt cactattaaa gtcagaaaat aaattctgtc gtagacaatg      240

ttaccataat gttcttgtcc attttgcata cactttaaat attcatttga tttctcaggg      300

ttcatgatca taataaattg cgcattcgca aggcggtagt attataatgg ggtccatcat      360

tctgtagcaa gaagttacag tacgctgttc aagcgttaaa caagataagt aatctcgaat      420

gaaacattca tatttcgcat gagccaacat acagttgctg agtaatcttc attgcgctta      480

tttatcggca ttgagattgt aaaggaagta aaacgcattt ttgcagatct gttctcttat      540

gtatttttaa tcgtccttgt atggaagtat caaaggggac gttcttcacc tccttggaac      600

gtatccctat ctggattaac atcactgcca cagatcgaat tgcaagaag                  649


<210>  76
<211>  674
<212>  DNA
<213>  artificial sequence

<220>
<223>  right flank RF sequence with overlap ScTAL1t - INT3 3' (part of 
       donor) - INT3 3' genomic DNA. Homology with INT3 3' genomic DNA 
       624 bp

<400>  76
gcagccccta agatcagaca aaacatccgg aaccacctta aatcaacgtc gatgggacgt       60

cagcactgta cttgtttttg cgactagatt gtaaatcatt ctttatttaa tctctttctt      120

taactactgc ttaaagtata atttggtccg tagtttaata actatactaa gcgtaacaat      180

gcatactgac attataagcc tgaacattac gagtttaagt tgtatgtagg cgttctgtaa      240

gaggttactg cgtaaattat caacgaatgc attggtgtat ttgcgaaagc tacttctttt      300

aacaagtatt tacataagaa taatggtgat ctgctcaact gatttggtga taactctaac      360

ttttttagca acaatttaaa agataattcg aacatatata acagtaggaa gaatttgtgt      420

acgtcaaatt aagataattt agcattacca aagttattaa cctaaacata aaatatatat      480

gagacacatg tggaaatcgt atgaaacaac tgttatgaaa ctgacaagaa tgaatatata      540

gagtaagctc cgcttgtaaa gaggaatcac ttaagtgtat aaatgtctcg acgattactt      600

tagatccaag attgatgatt gatattactc tgtaatactt aagctctttt aatagctcac      660

tgttgtatta cggg                                                        674


<210>  77
<211>  81
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer primer to obtain SEQ ID NO: 68

<400>  77
aattttgggg atattggctt ttttttttaa agtttacaaa tgaatttttt ccgccaggat       60

gaagtatgga tctttgaaac c                                                 81


<210>  78
<211>  24
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 68

<400>  78
cacaagctta ttcttccaaa aatc                                              24


<210>  79
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 69, 70 ,71

<400>  79
cattatatcg aggaaagccc                                                   20


<210>  80
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 69

<400>  80
cctactcctc tgcctccccg ccacatgggg gtgaccgcaa aaaaagaaaa cttgacacca       60

atgggctaac g                                                            71


<210>  81
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 70

<400>  81
ggcttgcggg tctttggcac catttatgag aaggcatttg cttggacagg cttgacacca       60

atgggctaac g                                                            71


<210>  82
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 71

<400>  82
cattgtattt tgaacgtaca aagtatgcat tgtttatacg ctattatcag ccttgacacc       60

aatgggctaa cg                                                           72


<210>  83
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 72

<400>  83
gaattagtgg aaccaaggaa aaaaaaagag gtatccttga ttaaggaaca gaacaatact       60

gattgcgtta cc                                                           72


<210>  84
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 72

<400>  84
cgcacaccac gaggtgaagg                                                   20


<210>  85
<211>  23
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 73, 74, 75

<400>  85
taaaggaggt gcacgcatta tgg                                               23


<210>  86
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 73

<400>  86
gttttttatt ttctgaaagg ttcaaagtat gcagaacaaa ttaatgtttg ttccaaggag       60

gtgaagaacg tc                                                           72


<210>  87
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 74

<400>  87
ggtagtatct gtcctcttgt tattgttact gtaattgtgt atatatgttt tccaaggagg       60

tgaagaacgt c                                                            71


<210>  88
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 75

<400>  88
cttcttgcaa ttcgatctgt ggcagtgatg ttaatccaga tagggatacg ttccaaggag       60

gtgaagaacg tc                                                           72


<210>  89
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain SEQ ID NO: 76

<400>  89
gcagccccta agatcagaca aaacatccgg aaccacctta aatcaacgtc gatgggacgt       60

cagcactgta c                                                            71


<210>  90
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain SEQ ID NO: 76

<400>  90
cccgtaatac aacagtgagc                                                   20


<210>  91
<211>  5712
<212>  DNA
<213>  artificial sequence

<220>
<223>  vector pRN1120

<400>  91
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca       60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg      120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc      180

accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc      240

accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca      300

ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat      360

taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc      420

ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc      480

aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt      540

agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg      600

tttctattat gaatttcatt tataaagttt atgtacacct aggatccgtc gacactggat      660

ggcggcgtta gtatcgaatc gacagcagta tagcgaccag cattcacata cgattgacgc      720

atgatattac tttctgcgca cttaacttcg catctgggca gatgatgtcg aggcgaaaaa      780

aaatataaat cacgctaaca tttgattaaa atagaacaac tacaatataa aaaaactata      840

caaatgacaa gttcttgaaa acaagaatct ttttattgtc agtactaggg gcagggcatg      900

ctcatgtaga gcgcctgctc gccgtccgag gcggtgccgt cgtacagggc ggtgtccagg      960

ccgcagaggg tgaaccccat ccgccggtac gcgtggatcg ccggtgcgtt gacgttggtg     1020

acctccagcc agaggtgccc ggcgccccgc tcgcgggcga actccgtcgc gagccccatc     1080

aacgcgcgcc cgaccccgtg cccccggtgc tccggggcga cctcgatgtc ctcgacggtc     1140

agccggcggt tccagccgga gtacgagacg accacgaagc ccgccaggtc gccgtcgtcc     1200

ccgtacgcga cgaacgtccg ggagtccggg tcgccgtcct ccccggcgtc cgattcgtcg     1260

tccgattcgt cgtcggggaa caccttggtc aggggcgggt ccaccggcac ctcccgcagg     1320

gtgaagccgt ccccggtggc ggtgacgcgg aagacggtgt cggtggtgaa ggacccatcc     1380

agtgcctcga tggcctcggc gtcccccggg acactggtgc ggtaccggta agccgtgtcg     1440

tcaagagtgg tcattttaca tggttgttta tgttcggatg tgatgtgaga actgtatcct     1500

agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaatc ctaacctttt     1560

atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag gggagcgttt     1620

ccctgctcgc aggtctgcag cgaggagccg taatttttgc ttcgcgccgt gcggccatca     1680

aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta cgggcgacag     1740

tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc     1800

atgtcgctgg ccgggtgacc cggcggggac gaggccttaa gttcgaacgt acgagctccg     1860

gcattgcgaa taccgctttc cacaaacatt gctcaaaagt atctctttgc tatatatctc     1920

tgtgctatat ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa     1980

ctcgacctct acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta     2040

gtaagaacta ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg     2100

tagtatatag agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat     2160

caacgctatc actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc     2220

ggggatgcct ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg     2280

gaagtggagt caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta     2340

accttaacgg acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag     2400

gagaaaaaaa gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt     2460

gtagaacaaa aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt     2520

ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca     2580

tttttgtttt acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg     2640

catttctgtt ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg     2700

ttgcattttt gttctacaaa atgaagcaca gatgcttcgt taacaaagat atgctattga     2760

agtgcaagat ggaaacgcag aaaatgaacc ggggatgcga cgtgcaagat tacctatgca     2820

atagatgcaa tagtttctcc aggaaccgaa atacatacat tgtcttccgt aaagcgctag     2880

actatatatt attatacagg ttcaaatata ctatctgttt cagggaaaac tcccaggttc     2940

ggatgttcaa aattcaatga tgggtaacaa gtacgatcgt aaatctgtaa aacagtttgt     3000

cggatattag gctgtatctc ctcaaagcgt attcgaatat cattgagaag ctgcagcgtc     3060

acatcggata ataatgatgg cagccattgt agaagtgcct tttgcatttc tagtctcttt     3120

ctcggtctag ctagttttac tacatcgcga agatagaatc ttagatcaca ctgcctttgc     3180

tgagctggat caatagagta acaaaagagt ggtaaggcct cgttaaagga caaggacctg     3240

agcggaagtg tatcgtacag tagacggagt atactaggta tagtctatag tccgtggaat     3300

taattctcat gtttgacagc ttatcatcga taatccggag ctagcatgcg gccgctctag     3360

aactagtgga tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct     3420

cgaggggggg cccggtaccc agcttttgtt ccctttagtg agggttaatt ccgagcttgg     3480

cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca     3540

acataggagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca     3600

cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc     3660

attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt     3720

cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact     3780

caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag     3840

caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata     3900

ggctcggccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc     3960

cgacaggact ataaagatac caggcgttcc cccctggaag ctccctcgtg cgctctcctg     4020

ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc     4080

tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg     4140

gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc     4200

ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga     4260

ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg     4320

gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa     4380

aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg     4440

tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt     4500

ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat     4560

tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct     4620

aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta     4680

tctcagcgat ctgtctattt cgttcatcca tagttgcctg actgcccgtc gtgtagataa     4740

ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac     4800

gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa     4860

gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag     4920

taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg     4980

tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag     5040

ttacatgatc ccccatgttg tgaaaaaaag cggttagctc cttcggtcct ccgatcgttg     5100

tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc     5160

ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat     5220

tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata     5280

ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa     5340

aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca     5400

actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc     5460

aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc     5520

tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg     5580

aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac     5640

ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga     5700

ggccctttcg tc                                                         5712


<210>  92
<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to remove SapI restriction site in pRN1120

<400>  92
tattgggccc tcttccgctt c                                                 21


<210>  93
<211>  18
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to remove SapI restriction site in pRN1120

<400>  93
cgcaaaccgc ctctcccc                                                     18


<210>  94
<211>  547
<212>  DNA
<213>  artificial sequence

<220>
<223>  gBlock allowing direct SapI cloning of the guide sequence, part 
       of vector pGRN002

<400>  94
ggagctagca tgcggccgct ctagaactag tggatccccc gggctgcagg aattctcttt       60

gaaaagataa tgtatgatta tgctttcact catatttata cagaaacttg atgttttctt      120

tcgagtatat acaaggtgat tacatgtacg tttgaagtac aactctagat tttgtagtgc      180

cctcttgggc tagcggtaaa ggtgcgcatt ttttcacacc ctacaatgtt ctgttcaaaa      240

gattttggtc aaacgctgta gaagtgaaag ttggtgcgca tgtttcggcg ttcgaaactt      300

ctccgcagtg aaagataaat gatcagaaga gcctgaggtc gacggtatcg ataagcttga      360

tatcaattcc ccgggggatc cactgctctt ctgttttaga gctagaaata gcaagttaaa      420

ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtggtg ctttttttgt      480

tttttatgtc tctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta      540

attccga                                                                547


<210>  95
<211>  6121
<212>  DNA
<213>  artificial sequence

<220>
<223>  expression vector pGRN002

<400>  95
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca       60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg      120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc      180

accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc      240

accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca      300

ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat      360

taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc      420

ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc      480

aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt      540

agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg      600

tttctattat gaatttcatt tataaagttt atgtacacct aggatccgtc gacactggat      660

ggcggcgtta gtatcgaatc gacagcagta tagcgaccag cattcacata cgattgacgc      720

atgatattac tttctgcgca cttaacttcg catctgggca gatgatgtcg aggcgaaaaa      780

aaatataaat cacgctaaca tttgattaaa atagaacaac tacaatataa aaaaactata      840

caaatgacaa gttcttgaaa acaagaatct ttttattgtc agtactaggg gcagggcatg      900

ctcatgtaga gcgcctgctc gccgtccgag gcggtgccgt cgtacagggc ggtgtccagg      960

ccgcagaggg tgaaccccat ccgccggtac gcgtggatcg ccggtgcgtt gacgttggtg     1020

acctccagcc agaggtgccc ggcgccccgc tcgcgggcga actccgtcgc gagccccatc     1080

aacgcgcgcc cgaccccgtg cccccggtgc tccggggcga cctcgatgtc ctcgacggtc     1140

agccggcggt tccagccgga gtacgagacg accacgaagc ccgccaggtc gccgtcgtcc     1200

ccgtacgcga cgaacgtccg ggagtccggg tcgccgtcct ccccggcgtc cgattcgtcg     1260

tccgattcgt cgtcggggaa caccttggtc aggggcgggt ccaccggcac ctcccgcagg     1320

gtgaagccgt ccccggtggc ggtgacgcgg aagacggtgt cggtggtgaa ggacccatcc     1380

agtgcctcga tggcctcggc gtcccccggg acactggtgc ggtaccggta agccgtgtcg     1440

tcaagagtgg tcattttaca tggttgttta tgttcggatg tgatgtgaga actgtatcct     1500

agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaatc ctaacctttt     1560

atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag gggagcgttt     1620

ccctgctcgc aggtctgcag cgaggagccg taatttttgc ttcgcgccgt gcggccatca     1680

aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta cgggcgacag     1740

tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc     1800

atgtcgctgg ccgggtgacc cggcggggac gaggccttaa gttcgaacgt acgagctccg     1860

gcattgcgaa taccgctttc cacaaacatt gctcaaaagt atctctttgc tatatatctc     1920

tgtgctatat ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa     1980

ctcgacctct acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta     2040

gtaagaacta ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg     2100

tagtatatag agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat     2160

caacgctatc actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc     2220

ggggatgcct ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg     2280

gaagtggagt caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta     2340

accttaacgg acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag     2400

gagaaaaaaa gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt     2460

gtagaacaaa aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt     2520

ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca     2580

tttttgtttt acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg     2640

catttctgtt ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg     2700

ttgcattttt gttctacaaa atgaagcaca gatgcttcgt taacaaagat atgctattga     2760

agtgcaagat ggaaacgcag aaaatgaacc ggggatgcga cgtgcaagat tacctatgca     2820

atagatgcaa tagtttctcc aggaaccgaa atacatacat tgtcttccgt aaagcgctag     2880

actatatatt attatacagg ttcaaatata ctatctgttt cagggaaaac tcccaggttc     2940

ggatgttcaa aattcaatga tgggtaacaa gtacgatcgt aaatctgtaa aacagtttgt     3000

cggatattag gctgtatctc ctcaaagcgt attcgaatat cattgagaag ctgcagcgtc     3060

acatcggata ataatgatgg cagccattgt agaagtgcct tttgcatttc tagtctcttt     3120

ctcggtctag ctagttttac tacatcgcga agatagaatc ttagatcaca ctgcctttgc     3180

tgagctggat caatagagta acaaaagagt ggtaaggcct cgttaaagga caaggacctg     3240

agcggaagtg tatcgtacag tagacggagt atactaggta tagtctatag tccgtggaat     3300

taattctcat gtttgacagc ttatcatcga taatccggag ctagcatgcg gccgctctag     3360

aactagtgga tcccccgggc tgcaggaatt ctctttgaaa agataatgta tgattatgct     3420

ttcactcata tttatacaga aacttgatgt tttctttcga gtatatacaa ggtgattaca     3480

tgtacgtttg aagtacaact ctagattttg tagtgccctc ttgggctagc ggtaaaggtg     3540

cgcatttttt cacaccctac aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag     3600

tgaaagttgg tgcgcatgtt tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc     3660

agaagagcct gaggtcgacg gtatcgataa gcttgatatc aattccccgg gggatccact     3720

gctcttctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt     3780

gaaaaagtgg caccgagtcg gtggtgcttt ttttgttttt tatgtctctc gagggggggc     3840

ccggtaccca gcttttgttc cctttagtga gggttaattc cgagcttggc gtaatcatgg     3900

tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa cataggagcc     3960

ggaagcataa agtgtaaagc ctggggtgcc taatgagtga ggtaactcac attaattgcg     4020

ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc     4080

ggccaacgcg cggggagagg cggtttgcgt attgggccct cttccgcttc ctcgctcact     4140

gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta     4200

atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag     4260

caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctcggcccc     4320

cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta     4380

taaagatacc aggcgttccc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg     4440

ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc     4500

tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac     4560

gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac     4620

ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg     4680

aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga     4740

aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt     4800

agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag     4860

cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct     4920

gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg     4980

atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat     5040

gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc     5100

tgtctatttc gttcatccat agttgcctga ctgcccgtcg tgtagataac tacgatacgg     5160

gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct     5220

ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca     5280

actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg     5340

ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg     5400

tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc     5460

cccatgttgt gaaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag     5520

ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg     5580

ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag     5640

tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat     5700

agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg     5760

atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca     5820

gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca     5880

aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat     5940

tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag     6000

aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa     6060

gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt     6120

c                                                                     6121


<210>  96
<211>  53
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer approach 1: 30 bp on either side

<400>  96
ataaatgatc tattagaacc agggaggtcc gttttagagc tagaaatagc aag              53


<210>  97
<211>  52
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer approach 1: 30 bp on either side

<400>  97
gctctaaaac ggacctccct ggttctaata gatcatttat ctttcactgc gg               52


<210>  98
<211>  6113
<212>  DNA
<213>  artificial sequence

<220>
<223>  PCR fragment approach 1: 30 bp on eiter side. The fragment was 
       transformed to yeast allowing reconstitution of a circular vector
       by in vivo recombination

<400>  98
ataaatgatc tattagaacc agggaggtcc gttttagagc tagaaatagc aagttaaaat       60

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct ttttttgttt      120

tttatgtctc tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat      180

tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac      240

aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt      300

gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc      360

gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcc      420

ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt      480

atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa      540

gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc      600

gtttttccat aggctcggcc cccctgacga gcatcacaaa aatcgacgct caagtcagag      660

gtggcgaaac ccgacaggac tataaagata ccaggcgttc ccccctggaa gctccctcgt      720

gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg      780

aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg      840

ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg      900

taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac      960

tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg     1020

gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt     1080

taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg     1140

tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc     1200

tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt     1260

ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt     1320

taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag     1380

tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactgcccgt     1440

cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc     1500

gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc     1560

cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg     1620

ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac     1680

aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg     1740

atcaaggcga gttacatgat cccccatgtt gtgaaaaaaa gcggttagct ccttcggtcc     1800

tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact     1860

gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc     1920

aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat     1980

acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc     2040

ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac     2100

tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa     2160

aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact     2220

catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg     2280

atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg     2340

aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag     2400

gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca     2460

catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc     2520

ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc     2580

agagcagatt gtactgagag tgcaccatat cgactacgtc gtaaggccgt ttctgacaga     2640

gtaaaattct tgagggaact ttcaccatta tgggaaatgg ttcaagaagg tattgactta     2700

aactccatca aatggtcagg tcattgagtg ttttttattt gttgtatttt ttttttttta     2760

gagaaaatcc tccaatatca aattaggaat cgtagtttca tgattttctg ttacacctaa     2820

ctttttgtgt ggtgccctcc tccttgtcaa tattaatgtt aaagtgcaat tctttttcct     2880

tatcacgttg agccattagt atcaatttgc ttacctgtat tcctttacta tcctcctttt     2940

tctccttctt gataaatgta tgtagattgc gtatatagtt tcgtctaccc tatgaacata     3000

ttccattttg taatttcgtg tcgtttctat tatgaatttc atttataaag tttatgtaca     3060

cctaggatcc gtcgacactg gatggcggcg ttagtatcga atcgacagca gtatagcgac     3120

cagcattcac atacgattga cgcatgatat tactttctgc gcacttaact tcgcatctgg     3180

gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta acatttgatt aaaatagaac     3240

aactacaata taaaaaaact atacaaatga caagttcttg aaaacaagaa tctttttatt     3300

gtcagtacta ggggcagggc atgctcatgt agagcgcctg ctcgccgtcc gaggcggtgc     3360

cgtcgtacag ggcggtgtcc aggccgcaga gggtgaaccc catccgccgg tacgcgtgga     3420

tcgccggtgc gttgacgttg gtgacctcca gccagaggtg cccggcgccc cgctcgcggg     3480

cgaactccgt cgcgagcccc atcaacgcgc gcccgacccc gtgcccccgg tgctccgggg     3540

cgacctcgat gtcctcgacg gtcagccggc ggttccagcc ggagtacgag acgaccacga     3600

agcccgccag gtcgccgtcg tccccgtacg cgacgaacgt ccgggagtcc gggtcgccgt     3660

cctccccggc gtccgattcg tcgtccgatt cgtcgtcggg gaacaccttg gtcaggggcg     3720

ggtccaccgg cacctcccgc agggtgaagc cgtccccggt ggcggtgacg cggaagacgg     3780

tgtcggtggt gaaggaccca tccagtgcct cgatggcctc ggcgtccccc gggacactgg     3840

tgcggtaccg gtaagccgtg tcgtcaagag tggtcatttt acatggttgt ttatgttcgg     3900

atgtgatgtg agaactgtat cctagcaaga ttttaaaagg aagtatatga aagaagaacc     3960

tcagtggcaa atcctaacct tttatatttc tctacagggg cgcggcgtgg ggacaattca     4020

acgcgtctgt gaggggagcg tttccctgct cgcaggtctg cagcgaggag ccgtaatttt     4080

tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa tgattataca tggggatgta     4140

tgggctaaat gtacgggcga cagtcacatc atgcccctga gctgcgcacg tcaagactgt     4200

caaggagggt attctgggcc tccatgtcgc tggccgggtg acccggcggg gacgaggcct     4260

taagttcgaa cgtacgagct ccggcattgc gaataccgct ttccacaaac attgctcaaa     4320

agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt     4380

cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt     4440

actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg     4500

aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa     4560

ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat     4620

ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct     4680

tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata     4740

gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag     4800

actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa     4860

tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa     4920

aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga     4980

aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt     5040

ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg     5100

tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt     5160

cgttaacaaa gatatgctat tgaagtgcaa gatggaaacg cagaaaatga accggggatg     5220

cgacgtgcaa gattacctat gcaatagatg caatagtttc tccaggaacc gaaatacata     5280

cattgtcttc cgtaaagcgc tagactatat attattatac aggttcaaat atactatctg     5340

tttcagggaa aactcccagg ttcggatgtt caaaattcaa tgatgggtaa caagtacgat     5400

cgtaaatctg taaaacagtt tgtcggatat taggctgtat ctcctcaaag cgtattcgaa     5460

tatcattgag aagctgcagc gtcacatcgg ataataatga tggcagccat tgtagaagtg     5520

ccttttgcat ttctagtctc tttctcggtc tagctagttt tactacatcg cgaagataga     5580

atcttagatc acactgcctt tgctgagctg gatcaataga gtaacaaaag agtggtaagg     5640

cctcgttaaa ggacaaggac ctgagcggaa gtgtatcgta cagtagacgg agtatactag     5700

gtatagtcta tagtccgtgg aattaattct catgtttgac agcttatcat cgataatccg     5760

gagctagcat gcggccgctc tagaactagt ggatcccccg ggctgcagga attctctttg     5820

aaaagataat gtatgattat gctttcactc atatttatac agaaacttga tgttttcttt     5880

cgagtatata caaggtgatt acatgtacgt ttgaagtaca actctagatt ttgtagtgcc     5940

ctcttgggct agcggtaaag gtgcgcattt tttcacaccc tacaatgttc tgttcaaaag     6000

attttggtca aacgctgtag aagtgaaagt tggtgcgcat gtttcggcgt tcgaaacttc     6060

tccgcagtga aagataaatg atctattaga accagggagg tccgttttag agc            6113


<210>  99
<211>  73
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer approach 1: 50 bp tail on one side

<400>  99
aaacttctcc gcagtgaaag ataaatgatc tattagaacc agggaggtcc gttttagagc       60

tagaaatagc aag                                                          73


<210>  100
<211>  22
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer approach 1: 50 bp tail on one side

<400>  100
gatcatttat ctttcactgc gg                                                22


<210>  101
<211>  6103
<212>  DNA
<213>  artificial sequence

<220>
<223>  PCR fragment approach 2: 50 bp tail on one side. The fragment was
       transformed to yeast allowing reconstitution of a circular vector
       by in vivo recombination

<400>  101
aaacttctcc gcagtgaaag ataaatgatc tattagaacc agggaggtcc gttttagagc       60

tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt      120

cggtggtgct ttttttgttt tttatgtctc tcgagggggg gcccggtacc cagcttttgt      180

tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg      240

tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat aaagtgtaaa      300

gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct      360

ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga      420

ggcggtttgc gtattgggcc ctcttccgct tcctcgctca ctgactcgct gcgctcggtc      480

gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa      540

tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt      600

aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga gcatcacaaa      660

aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttc      720

ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg      780

tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc      840

agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc      900

gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta      960

tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct     1020

acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc     1080

tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa     1140

caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa     1200

aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa     1260

aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt     1320

ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac     1380

agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc     1440

atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt accatctggc     1500

cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata     1560

aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc     1620

cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc     1680

aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca     1740

ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgaaaaaaa     1800

gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca     1860

ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt     1920

tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt     1980

tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg     2040

ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga     2100

tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc     2160

agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg     2220

acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag     2280

ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg     2340

gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg     2400

acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat     2460

gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg     2520

gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc     2580

tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat cgactacgtc     2640

gtaaggccgt ttctgacaga gtaaaattct tgagggaact ttcaccatta tgggaaatgg     2700

ttcaagaagg tattgactta aactccatca aatggtcagg tcattgagtg ttttttattt     2760

gttgtatttt ttttttttta gagaaaatcc tccaatatca aattaggaat cgtagtttca     2820

tgattttctg ttacacctaa ctttttgtgt ggtgccctcc tccttgtcaa tattaatgtt     2880

aaagtgcaat tctttttcct tatcacgttg agccattagt atcaatttgc ttacctgtat     2940

tcctttacta tcctcctttt tctccttctt gataaatgta tgtagattgc gtatatagtt     3000

tcgtctaccc tatgaacata ttccattttg taatttcgtg tcgtttctat tatgaatttc     3060

atttataaag tttatgtaca cctaggatcc gtcgacactg gatggcggcg ttagtatcga     3120

atcgacagca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc     3180

gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta     3240

acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg     3300

aaaacaagaa tctttttatt gtcagtacta ggggcagggc atgctcatgt agagcgcctg     3360

ctcgccgtcc gaggcggtgc cgtcgtacag ggcggtgtcc aggccgcaga gggtgaaccc     3420

catccgccgg tacgcgtgga tcgccggtgc gttgacgttg gtgacctcca gccagaggtg     3480

cccggcgccc cgctcgcggg cgaactccgt cgcgagcccc atcaacgcgc gcccgacccc     3540

gtgcccccgg tgctccgggg cgacctcgat gtcctcgacg gtcagccggc ggttccagcc     3600

ggagtacgag acgaccacga agcccgccag gtcgccgtcg tccccgtacg cgacgaacgt     3660

ccgggagtcc gggtcgccgt cctccccggc gtccgattcg tcgtccgatt cgtcgtcggg     3720

gaacaccttg gtcaggggcg ggtccaccgg cacctcccgc agggtgaagc cgtccccggt     3780

ggcggtgacg cggaagacgg tgtcggtggt gaaggaccca tccagtgcct cgatggcctc     3840

ggcgtccccc gggacactgg tgcggtaccg gtaagccgtg tcgtcaagag tggtcatttt     3900

acatggttgt ttatgttcgg atgtgatgtg agaactgtat cctagcaaga ttttaaaagg     3960

aagtatatga aagaagaacc tcagtggcaa atcctaacct tttatatttc tctacagggg     4020

cgcggcgtgg ggacaattca acgcgtctgt gaggggagcg tttccctgct cgcaggtctg     4080

cagcgaggag ccgtaatttt tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa     4140

tgattataca tggggatgta tgggctaaat gtacgggcga cagtcacatc atgcccctga     4200

gctgcgcacg tcaagactgt caaggagggt attctgggcc tccatgtcgc tggccgggtg     4260

acccggcggg gacgaggcct taagttcgaa cgtacgagct ccggcattgc gaataccgct     4320

ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata     4380

taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt     4440

ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag     4500

agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa     4560

atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct     4620

gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt     4680

gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt     4740

tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca     4800

gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct     4860

aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag     4920

tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc     4980

agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa     5040

tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa     5100

aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac     5160

aaaatgaagc acagatgctt cgttaacaaa gatatgctat tgaagtgcaa gatggaaacg     5220

cagaaaatga accggggatg cgacgtgcaa gattacctat gcaatagatg caatagtttc     5280

tccaggaacc gaaatacata cattgtcttc cgtaaagcgc tagactatat attattatac     5340

aggttcaaat atactatctg tttcagggaa aactcccagg ttcggatgtt caaaattcaa     5400

tgatgggtaa caagtacgat cgtaaatctg taaaacagtt tgtcggatat taggctgtat     5460

ctcctcaaag cgtattcgaa tatcattgag aagctgcagc gtcacatcgg ataataatga     5520

tggcagccat tgtagaagtg ccttttgcat ttctagtctc tttctcggtc tagctagttt     5580

tactacatcg cgaagataga atcttagatc acactgcctt tgctgagctg gatcaataga     5640

gtaacaaaag agtggtaagg cctcgttaaa ggacaaggac ctgagcggaa gtgtatcgta     5700

cagtagacgg agtatactag gtatagtcta tagtccgtgg aattaattct catgtttgac     5760

agcttatcat cgataatccg gagctagcat gcggccgctc tagaactagt ggatcccccg     5820

ggctgcagga attctctttg aaaagataat gtatgattat gctttcactc atatttatac     5880

agaaacttga tgttttcttt cgagtatata caaggtgatt acatgtacgt ttgaagtaca     5940

actctagatt ttgtagtgcc ctcttgggct agcggtaaag gtgcgcattt tttcacaccc     6000

tacaatgttc tgttcaaaag attttggtca aacgctgtag aagtgaaagt tggtgcgcat     6060

gtttcggcgt tcgaaacttc tccgcagtga aagataaatg atc                       6103


<210>  102
<211>  23
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain the 5' flank A - connector 5 sequence to
       obtain 1 kB deletion of genomic DNA

<400>  102
cactatagca atctggctat atg                                               23


<210>  103
<211>  24
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain the 5' flank A and B - connector 5 
       sequence to obtain 1 kB deletion of genomic DNA

<400>  103
aaacgcctgt gggtgtggta ctgg                                              24


<210>  104
<211>  500
<212>  DNA
<213>  artificial sequence

<220>
<223>  fragment A (5' flank A - connector 5) to obtain 1 kB deletion of 
       genomic DNA

<400>  104
cactatagca atctggctat atgatatgca gaaaatagtg attactgtgc tctttgctgc       60

atcatgcatc atatgtttgg cattctgtcc tgtcagatga atgggcgaaa caattcgggg      120

agctttgttg cgacttggta cccggcaagc cgcgacctat attttgcatt aaacgagttc      180

atcgtggatt tatcgccatg agccttagct tatcaagcct ctcacagaca attaagcaat      240

gaaaaaggta taccatcggc gcagaatggt taactagtgg gttcatactg ctgtgttata      300

gattgttacc taagtgatca ccaaaaaaaa gtgcaaaaag gaaaaaaaaa taagagacag      360

gtaacttcca caagcttatt cttccaaaaa tcaatcttat cttcatgcca gcaatagttg      420

cgtgctgagc tcaacagtgc ccaacccttg aagcgacttc caatcgcttt gcatatccag      480

taccacaccc acaggcgttt                                                  500


<210>  105
<211>  26
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain the 5' flank B - connector 5 sequence to
       obtain 10 kB deletion of genomic DNA

<400>  105
caatagtaat tttaaaaaca atgtgc                                            26


<210>  106
<211>  500
<212>  DNA
<213>  artificial sequence

<220>
<223>  fragment B (5' flank B - connector 5) to obtain 10 kB deletion of
       genomic DNA

<400>  106
caatagtaat tttaaaaaca atgtgcttat ccagaaattc cataaggagt agcgcttgga       60

tccctcacct ttttcggtcg cggcagtctt caaaaccttt tttggctcgt catttttttt      120

taactttgcc ttctttgtca cagccatatt ttgcctcgaa cgatcacctt tcatcgtagg      180

ctatccaata gacgtcaaat gccttgctga ccactacaga actgtaatga acttcagcta      240

tttttaacgt atgttattct ataattctat accactattg tcgttttgaa aaatgaaatt      300

tcacttttcg gaagttgtcc gctaagaaca actaagtgat aacagtaaat tttaaaaatg      360

ctttgaaaaa tgggttaagc aaaggttatt tactataact acgtagtgaa cgaaaccata      420

tatctcttaa tttgcagcag ggattttata aagcgacttc caatcgcttt gcatatccag      480

taccacaccc acaggcgttt                                                  500


<210>  107
<211>  22
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain Connector 5 - TDH3p - GFP - ENO1t - 
       Connector 3 PCR fragment

<400>  107
aagcgacttc caatcgcttt gc                                                22


<210>  108
<211>  24
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain Connector 5 - TDH3p - GFP - ENO1t - 
       Connector 3 PCR fragment

<400>  108
acttagtatg gtctgttgga aagg                                              24


<210>  109
<211>  1726
<212>  DNA
<213>  artificial sequence

<220>
<223>  fragment C, Connector 5 - TDH3p - GFP - ENO1t - Connector 3 
       synthetic cassette

<400>  109
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt gtgcttagtc       60

aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta      120

cacagaatat ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa      180

tataatggag cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa      240

tattgttttc ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga      300

acaggggcac aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg      360

gagtaaatga tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc      420

ttctattacc ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt      480

tccctgaaat tattccccta cttgactaat aagtatataa agacggtagg tattgattgt      540

aattctgtaa atctatttct taaacttctt aaattctact tttatagtta gtcttttttt      600

tagttttaaa acaccaagaa cttagtttcg aataaacaca cataaacaaa caaaatgtct      660

aaaggtgaag aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt      720

aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg      780

accttaaaat tgatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact      840

actttaggtt atggtttgca atgttttgct agatacccag atcatatgaa acaacatgac      900

tttttcaagt ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat      960

gacggtaact acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga     1020

atcgaattaa aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa     1080

tacaactata actctcacaa tgtttacatc actgctgaca aacaaaagaa tggtatcaaa     1140

gctaacttca aaattagaca caacattgaa gatggtggtg ttcaattagc tgaccattat     1200

caacaaaata ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc     1260

tatcaatctg ccttatccaa agatccaaac gaaaagagag atcacatggt cttgttagaa     1320

tttgttactg ctgctggtat tacccatggt atggatgaat tgtacaaata aaagcttttg     1380

attaagcctt ctagtccaaa aaacacgttt ttttgtcatt tatttcattt tcttagaata     1440

gtttagttta ttcattttat agtcacgaat gttttatgat tctatatagg gttgcaaaca     1500

agcatttttc attttatgtt aaaacaattt caggtttacc ttttattctg cttgtggtga     1560

cgcgtgtatc cgcccgctct tttggtcacc catgtattta attgcataaa taattcttaa     1620

aagtggagct agtctatttc tatttacata cctctcattt ctcatttcct cccctcagaa     1680

agcctgtatg cgaagccaca atcctttcca acagaccata ctaagt                    1726


<210>  110
<211>  72
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain the connector 3 - 3' flank A sequence to
       obtain 1 kB deletion of genomic DNA

<400>  110
agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt aagcgttgaa       60

gtttcctctt tg                                                           72


<210>  111
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain the connector 3 - 3' flank A and B 
       sequence to obtain 1 kB deletion of genomic DNA

<400>  111
tgtcaactgg agagctatcg                                                   20


<210>  112
<211>  581
<212>  DNA
<213>  artificial sequence

<220>
<223>  fragment D (connector 3 - 3' flank A) to obtain 1 kB deletion of 
       genomic DNA

<400>  112
agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt aagcgttgaa       60

gtttcctctt tgtatatttg agatcttcat tttatcggat tctttgtcat cagacaactt      120

gttgagtggt actaaaggag tgcttttcat catccttttg gtgaacgatt tcaaatacgt      180

tagtgttttc tgagctagtt ttgatcaatt caggtgattc gttatcagaa ctctcaggtt      240

tgtattcgtg tccagttgtg tagcattcgc ctaacgtgta agcacggatt tcttcctcag      300

aaatttcact gtatggaatc atgcccttct ttctcgcttc ttcgtcggta aatgcaccat      360

agtaatcttt gtcatcatgt ctaacagtaa ttttgaatgg gaagaagaca catagccccc      420

agtaaacgaa aaaagaaatc aaaaaggaga agaaagaatc accataaaag aatttaacaa      480

tacctgagtc gtggaaatag ttattgttga cttcccaagc gataccaggt agaccgggag      540

ccataccaca cacccaggca acgatagctc tccagttgac a                          581


<210>  113
<211>  71
<212>  DNA
<213>  artificial sequence

<220>
<223>  forward primer to obtain the connector 3 - 3' flank B sequence to
       obtain 10 kB deletion of genomic DNA

<400>  113
agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt ctgttgtgac       60

agcatcttgt c                                                            71


<210>  114
<211>  22
<212>  DNA
<213>  artificial sequence

<220>
<223>  reverse primer to obtain the connector 3 - 3' flank B sequence to
       obtain 10 kB deletion of genomic DNA

<400>  114
cctagcagta gttgtaatgt gg                                                22


<210>  115
<211>  607
<212>  DNA
<213>  artificial sequence

<220>
<223>  fragment E (connector 3 - 3' flank B) to obtain 10 kB deletion of
       genomic DNA

<400>  115
agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt ctgttgtgac       60

agcatcttgt ctattattta ttggatacgc aaaactcatt tttttaaaat tttttttata      120

atttccttta aataaaaagg ttaaagctgg gaaaaatgca tctaaataat aaactaatga      180

tttagtttcc ctcatttaaa atatctaaga aggacagaac tttgtttggg gtacttaccc      240

ttgagtattt ttagcgatta ttcttggagt tctcagccag aagattaaaa gaatatatgt      300

tactgaatgc agtgtgatac tcaagactaa gtagtcttaa caacaagtta acaagggcgt      360

gtggtctagt ggtatgattc tcgctttggg cgacttcctg actaacagga agacaaagca      420

tgcgagaggc cctgggttca attcccagct cgcccctttt atttttgatt atttatttcc      480

tattggtcgg gtaacagatt ttttttattt ttttcaaggc ccttttgttt tgtaaacaaa      540

gaaatttcac aaagtgagcg tctactaata ttgataatgc catttccaca ttacaactac      600

tgctagg                                                                607


<210>  116
<211>  15
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of the first 15 polynucleotides 
       (nucleotide 1-15) in the forward primers according to SEQ ID NO: 
       7 to 10

<400>  116
tgaaagataa atgat                                                        15


<210>  117
<211>  25
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of the last 25 polynucleotides 
       (nucleotide 36-60) in the forward primers according to SEQ ID NO:
       7 to 10

<400>  117
gttttagagc tagaaatagc aagtt                                             25


<210>  118
<211>  15
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of the first 15 polynucleotides 
       (nucleotide 1-15) in the reverse primers according to SEQ ID NO: 
       11 to 14

<400>  118
ttctagctct aaaac                                                        15


<210>  119
<211>  25
<212>  DNA
<213>  artificial sequence

<220>
<223>  polynucleotide sequence of the last 25 polynucleotides 
       (nucleotide 36-60) in the reverse primers according to SEQ ID NO:
       11 to 14

<400>  119
atcatttatc tttcactgcg gagaa                                             25


