                         SEQUENCE LISTING

<110>  E.I. DUPONT DE NEMOURS AND COMPANY and  
	 PIONEER HI-BRED INTERNATIONAL, INC.
       Cigan, Mark
       Patten, Phillip
       Young, Josh
 
<120>  GENOME MODIFICATION USING GUIDE P0LYNUCLEOTIDE/CAS ENDONUCLEASE 
       SYSTEMS AND METHODS OF USE

<130>  BB2344PCT

<150>  61/868706
<151>  2013-08-22

<150>  61/882532
<151>  2013-09-25

<150>  61/937045
<151>  2014-02-07

<150>  61/953090
<151>  2014-03-14

<150>  62/023239
<151>  2014-07-11

<160>  96    

<170>  PatentIn version 3.5

<210>  1
<211>  4107
<212>  DNA
<213>  Streptococcus pyogenes M1 GAS (SF370)

<400>  1
atggataaga aatactcaat aggcttagat atcggcacaa atagcgtcgg atgggcggtg       60

atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc      120

cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa      180

gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt      240

tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga      300

cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga      360

aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa      420

aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat      480

atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat      540

gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct      600

attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga      660

cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat      720

ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa      780

gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg      840

caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt      900

ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca      960

atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga     1020

caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca     1080

ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta     1140

gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc     1200

aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat     1260

gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt     1320

gaaaaaatct tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt     1380

cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa     1440

gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa     1500

aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt     1560

tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt     1620

tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc     1680

gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt     1740

tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt     1800

attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt     1860

ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct     1920

cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga     1980

cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta     2040

gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat     2100

agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta     2160

catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact     2220

gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt     2280

attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt     2340

atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct     2400

gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga     2460

gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatcac     2520

attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct     2580

gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa     2640

aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta     2700

acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa     2760

ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat     2820

actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct     2880

aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat     2940

taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa     3000

tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa     3060

atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct     3120

aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc     3180

cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt     3240

gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagta     3300

cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt     3360

gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct     3420

tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt     3480

aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac     3540

tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa     3600

tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta     3660

caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt     3720

cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag     3780

cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt     3840

attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa     3900

ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct     3960

cccgctgctt ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa     4020

gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt     4080

gatttgagtc agctaggagg tgactga                                         4107


<210>  2
<211>  189
<212>  DNA
<213>  Solanum tuberosum

<400>  2
gtaagtttct gcttctacct ttgatatata tataataatt atcattaatt agtagtaata       60

taatatttca aatatttttt tcaaaataaa agaatgtagt atatagcaat tgcttttctg      120

tagtttataa gtgtgtatat tttaatttat aacttttcta atatatgacc aaaacatggt      180

gatgtgcag                                                              189


<210>  3
<211>  9
<212>  PRT
<213>  Simian virus 40

<400>  3

Met Ala Pro Lys Lys Lys Arg Lys Val 
1               5                   


<210>  4
<211>  18
<212>  PRT
<213>  Agrobacterium tumefaciens

<400>  4

Lys Arg Pro Arg Asp Arg His Asp Gly Glu Leu Gly Gly Arg Lys Arg 
1               5                   10                  15      


Ala Arg 
        


<210>  5
<211>  6717
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Maize optimized Cas9 expression cassette

<400>  5
gtgcagcgtg acccggtcgt gcccctctct agagataatg agcattgcat gtctaagtta       60

taaaaaatta ccacatattt tttttgtcac acttgtttga agtgcagttt atctatcttt      120

atacatatat ttaaacttta ctctacgaat aatataatct atagtactac aataatatca      180

gtgttttaga gaatcatata aatgaacagt tagacatggt ctaaaggaca attgagtatt      240

ttgacaacag gactctacag ttttatcttt ttagtgtgca tgtgttctcc tttttttttg      300

caaatagctt cacctatata atacttcatc cattttatta gtacatccat ttagggttta      360

gggttaatgg tttttataga ctaatttttt tagtacatct attttattct attttagcct      420

ctaaattaag aaaactaaaa ctctatttta gtttttttat ttaataattt agatataaaa      480

tagaataaaa taaagtgact aaaaattaaa caaataccct ttaagaaatt aaaaaaacta      540

aggaaacatt tttcttgttt cgagtagata atgccagcct gttaaacgcc gtcgacgagt      600

ctaacggaca ccaaccagcg aaccagcagc gtcgcgtcgg gccaagcgaa gcagacggca      660

cggcatctct gtcgctgcct ctggacccct ctcgagagtt ccgctccacc gttggacttg      720

ctccgctgtc ggcatccaga aattgcgtgg cggagcggca gacgtgagcc ggcacggcag      780

gcggcctcct cctcctctca cggcaccggc agctacgggg gattcctttc ccaccgctcc      840

ttcgctttcc cttcctcgcc cgccgtaata aatagacacc ccctccacac cctctttccc      900

caacctcgtg ttgttcggag cgcacacaca cacaaccaga tctcccccaa atccacccgt      960

cggcacctcc gcttcaaggt acgccgctcg tcctcccccc cccccctctc taccttctct     1020

agatcggcgt tccggtccat gcatggttag ggcccggtag ttctacttct gttcatgttt     1080

gtgttagatc cgtgtttgtg ttagatccgt gctgctagcg ttcgtacacg gatgcgacct     1140

gtacgtcaga cacgttctga ttgctaactt gccagtgttt ctctttgggg aatcctggga     1200

tggctctagc cgttccgcag acgggatcga tttcatgatt ttttttgttt cgttgcatag     1260

ggtttggttt gcccttttcc tttatttcaa tatatgccgt gcacttgttt gtcgggtcat     1320

cttttcatgc ttttttttgt cttggttgtg atgatgtggt ctggttgggc ggtcgttcta     1380

gatcggagta gaattctgtt tcaaactacc tggtggattt attaattttg gatctgtatg     1440

tgtgtgccat acatattcat agttacgaat tgaagatgat ggatggaaat atcgatctag     1500

gataggtata catgttgatg cgggttttac tgatgcatat acagagatgc tttttgttcg     1560

cttggttgtg atgatgtggt gtggttgggc ggtcgttcat tcgttctaga tcggagtaga     1620

atactgtttc aaactacctg gtgtatttat taattttgga actgtatgtg tgtgtcatac     1680

atcttcatag ttacgagttt aagatggatg gaaatatcga tctaggatag gtatacatgt     1740

tgatgtgggt tttactgatg catatacatg atggcatatg cagcatctat tcatatgctc     1800

taaccttgag tacctatcta ttataataaa caagtatgtt ttataattat tttgatcttg     1860

atatacttgg atgatggcat atgcagcagc tatatgtgga tttttttagc cctgccttca     1920

tacgctattt atttgcttgg tactgtttct tttgtcgatg ctcaccctgt tgtttggtgt     1980

tacttctgca ggtcgactct agaggatcca tggcaccgaa gaagaagcgc aaggtgatgg     2040

acaagaagta cagcatcggc ctcgacatcg gcaccaactc ggtgggctgg gccgtcatca     2100

cggacgaata taaggtcccg tcgaagaagt tcaaggtcct cggcaataca gaccgccaca     2160

gcatcaagaa aaacttgatc ggcgccctcc tgttcgatag cggcgagacc gcggaggcga     2220

ccaggctcaa gaggaccgcc aggagacggt acactaggcg caagaacagg atctgctacc     2280

tgcaggagat cttcagcaac gagatggcga aggtggacga ctccttcttc caccgcctgg     2340

aggaatcatt cctggtggag gaggacaaga agcatgagcg gcacccaatc ttcggcaaca     2400

tcgtcgacga ggtaagtttc tgcttctacc tttgatatat atataataat tatcattaat     2460

tagtagtaat ataatatttc aaatattttt ttcaaaataa aagaatgtag tatatagcaa     2520

ttgcttttct gtagtttata agtgtgtata ttttaattta taacttttct aatatatgac     2580

caaaacatgg tgatgtgcag gtggcctacc acgagaagta cccgacaatc taccacctcc     2640

ggaagaaact ggtggacagc acagacaagg cggacctccg gctcatctac cttgccctcg     2700

cgcatatgat caagttccgc ggccacttcc tcatcgaggg cgacctgaac ccggacaact     2760

ccgacgtgga caagctgttc atccagctcg tgcagacgta caatcaactg ttcgaggaga     2820

accccataaa cgctagcggc gtggacgcca aggccatcct ctcggccagg ctctcgaaat     2880

caagaaggct ggagaacctt atcgcgcagt tgccaggcga aaagaagaac ggcctcttcg     2940

gcaaccttat tgcgctcagc ctcggcctga cgccgaactt caaatcaaac ttcgacctcg     3000

cggaggacgc caagctccag ctctcaaagg acacctacga cgacgacctc gacaacctcc     3060

tggcccagat aggagaccag tacgcggacc tcttcctcgc cgccaagaac ctctccgacg     3120

ctatcctgct cagcgacatc cttcgggtca acaccgaaat taccaaggca ccgctgtccg     3180

ccagcatgat taaacgctac gacgagcacc atcaggacct cacgctgctc aaggcactcg     3240

tccgccagca gctccccgag aagtacaagg agatcttctt cgaccaatca aaaaacggct     3300

acgcgggata tatcgacggc ggtgccagcc aggaagagtt ctacaagttc atcaaaccaa     3360

tcctggagaa gatggacggc accgaggagt tgctggtcaa gctcaacagg gaggacctcc     3420

tcaggaagca gaggaccttc gacaacggct ccatcccgca tcagatccac ctgggcgaac     3480

tgcatgccat cctgcggcgc caggaggact tctacccgtt cctgaaggat aaccgggaga     3540

agatcgagaa gatcttgacg ttccgcatcc catactacgt gggcccgctg gctcgcggca     3600

actcccggtt cgcctggatg acccggaagt cggaggagac catcacaccc tggaactttg     3660

aggaggtggt cgataagggc gctagcgctc agagcttcat cgagcgcatg accaacttcg     3720

ataaaaacct gcccaatgaa aaagtcctcc ccaagcactc gctgctctac gagtacttca     3780

ccgtgtacaa cgagctcacc aaggtcaaat acgtcaccga gggcatgcgg aagccggcgt     3840

tcctgagcgg cgagcagaag aaggcgatag tggacctcct cttcaagacc aacaggaagg     3900

tgaccgtgaa gcaattaaaa gaggactact tcaagaaaat agagtgcttc gactccgtgg     3960

agatctcggg cgtggaggat cggttcaacg cctcactcgg cacgtatcac gacctcctca     4020

agatcattaa agacaaggac ttcctcgaca acgaggagaa cgaggacatc ctcgaggaca     4080

tcgtcctcac cctgaccctg ttcgaggacc gcgaaatgat cgaggagagg ctgaagacct     4140

acgcgcacct gttcgacgac aaggtcatga aacagctcaa gaggcgccgc tacactggtt     4200

ggggaaggct gtcccgcaag ctcattaatg gcatcaggga caagcagagc ggcaagacca     4260

tcctggactt cctcaagtcc gacgggttcg ccaaccgcaa cttcatgcag ctcattcacg     4320

acgactcgct cacgttcaag gaagacatcc agaaggcaca ggtgagcggg cagggtgact     4380

ccctccacga acacatcgcc aacctggccg gctcgccggc cattaaaaag ggcatcctgc     4440

agacggtcaa ggtcgtcgac gagctcgtga aggtgatggg ccggcacaag cccgaaaata     4500

tcgtcataga gatggccagg gagaaccaga ccacccaaaa agggcagaag aactcgcgcg     4560

agcggatgaa acggatcgag gagggcatta aagagctcgg gtcccagatc ctgaaggagc     4620

accccgtgga aaatacccag ctccagaatg aaaagctcta cctctactac ctgcagaacg     4680

gccgcgacat gtacgtggac caggagctgg acattaatcg gctatcggac tacgacgtcg     4740

accacatcgt gccgcagtcg ttcctcaagg acgatagcat cgacaacaag gtgctcaccc     4800

ggtcggataa aaatcggggc aagagcgaca acgtgcccag cgaggaggtc gtgaagaaga     4860

tgaaaaacta ctggcgccag ctcctcaacg cgaaactgat cacccagcgc aagttcgaca     4920

acctgacgaa ggcggaacgc ggtggcttga gcgaactcga taaggcgggc ttcataaaaa     4980

ggcagctggt cgagacgcgc cagatcacga agcatgtcgc ccagatcctg gacagccgca     5040

tgaatactaa gtacgatgaa aacgacaagc tgatccggga ggtgaaggtg atcacgctga     5100

agtccaagct cgtgtcggac ttccgcaagg acttccagtt ctacaaggtc cgcgagatca     5160

acaactacca ccacgcccac gacgcctacc tgaatgcggt ggtcgggacc gccctgatca     5220

agaagtaccc gaagctggag tcggagttcg tgtacggcga ctacaaggtc tacgacgtgc     5280

gcaaaatgat cgccaagtcc gagcaggaga tcggcaaggc cacggcaaaa tacttcttct     5340

actcgaacat catgaacttc ttcaagaccg agatcaccct cgcgaacggc gagatccgca     5400

agcgcccgct catcgaaacc aacggcgaga cgggcgagat cgtctgggat aagggccggg     5460

atttcgcgac ggtccgcaag gtgctctcca tgccgcaagt caatatcgtg aaaaagacgg     5520

aggtccagac gggcgggttc agcaaggagt ccatcctccc gaagcgcaac tccgacaagc     5580

tcatcgcgag gaagaaggat tgggacccga aaaaatatgg cggcttcgac agcccgaccg     5640

tcgcatacag cgtcctcgtc gtggcgaagg tggagaaggg caagtcaaag aagctcaagt     5700

ccgtgaagga gctgctcggg atcacgatta tggagcggtc ctccttcgag aagaacccga     5760

tcgacttcct agaggccaag ggatataagg aggtcaagaa ggacctgatt attaaactgc     5820

cgaagtactc gctcttcgag ctggaaaacg gccgcaagag gatgctcgcc tccgcaggcg     5880

agttgcagaa gggcaacgag ctcgccctcc cgagcaaata cgtcaatttc ctgtacctcg     5940

ctagccacta tgaaaagctc aagggcagcc cggaggacaa cgagcagaag cagctcttcg     6000

tggagcagca caagcattac ctggacgaga tcatcgagca gatcagcgag ttctcgaagc     6060

gggtgatcct cgccgacgcg aacctggaca aggtgctgtc ggcatataac aagcaccgcg     6120

acaaaccaat acgcgagcag gccgaaaata tcatccacct cttcaccctc accaacctcg     6180

gcgctccggc agccttcaag tacttcgaca ccacgattga ccggaagcgg tacacgagca     6240

cgaaggaggt gctcgatgcg acgctgatcc accagagcat cacagggctc tatgaaacac     6300

gcatcgacct gagccagctg ggcggagaca agagaccacg ggaccgccac gatggcgagc     6360

tgggaggccg caagcgggca aggtaggtac cgttaaccta gacttgtcca tcttctggat     6420

tggccaactt aattaatgta tgaaataaaa ggatgcacac atagtgacat gctaatcact     6480

ataatgtggg catcaaagtt gtgtgttatg tgtaattact agttatctga ataaaagaga     6540

aagagatcat ccatatttct tatcctaaat gaatgtcacg tgtctttata attctttgat     6600

gaaccagatg catttcatta accaaatcca tatacatata aatattaatc atatataatt     6660

aatatcaatt gggttagcaa aacaaatcta gtctaggtgt gttttgcgaa tgcggcc        6717


<210>  6
<211>  1000
<212>  DNA
<213>  Zea mays

<400>  6
tgagagtaca atgatgaacc tagattaatc aatgccaaag tctgaaaaat gcaccctcag       60

tctatgatcc agaaaatcaa gattgcttga ggccctgttc ggttgttccg gattagagcc      120

ccggattaat tcctagccgg attacttctc taatttatat agattttgat gagctggaat      180

gaatcctggc ttattccggt acaaccgaac aggccctgaa ggataccagt aatcgctgag      240

ctaaattggc atgctgtcag agtgtcagta ttgcagcaag gtagtgagat aaccggcatc      300

atggtgccag tttgatggca ccattagggt tagagatggt ggccatgggc gcatgtcctg      360

gccaactttg tatgatatat ggcagggtga ataggaaagt aaaattgtat tgtaaaaagg      420

gatttcttct gtttgttagc gcatgtacaa ggaatgcaag ttttgagcga gggggcatca      480

aagatctggc tgtgtttcca gctgtttttg ttagccccat cgaatccttg acataatgat      540

cccgcttaaa taagcaacct cgcttgtata gttccttgtg ctctaacaca cgatgatgat      600

aagtcgtaaa atagtggtgt ccaaagaatt tccaggccca gttgtaaaag ctaaaatgct      660

attcgaattt ctactagcag taagtcgtgt ttagaaatta tttttttata tacctttttt      720

ccttctatgt acagtaggac acagtgtcag cgccgcgttg acggagaata tttgcaaaaa      780

agtaaaagag aaagtcatag cggcgtatgt gccaaaaact tcgtcacaga gagggccata      840

agaaacatgg cccacggccc aatacgaagc accgcgacga agcccaaaca gcagtccgta      900

ggtggagcaa agcgctgggt aatacgcaaa cgttttgtcc caccttgact aatcacaaga      960

gtggagcgta ccttataaac cgagccgcaa gcaccgaatt                           1000


<210>  7
<211>  11
<212>  PRT
<213>  artificial sequence

<220>
<223>  SV40 (Simian virus 40) Nuclear localization signal

<400>  7

Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val 
1               5                   10      


<210>  8
<211>  1047
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Maize optimized crRNA expression cassette containing the LIGCas-3
       target sequence in the variable targeting domain

<400>  8
tgagagtaca atgatgaacc tagattaatc aatgccaaag tctgaaaaat gcaccctcag       60

tctatgatcc agaaaatcaa gattgcttga ggccctgttc ggttgttccg gattagagcc      120

ccggattaat tcctagccgg attacttctc taatttatat agattttgat gagctggaat      180

gaatcctggc ttattccggt acaaccgaac aggccctgaa ggataccagt aatcgctgag      240

ctaaattggc atgctgtcag agtgtcagta ttgcagcaag gtagtgagat aaccggcatc      300

atggtgccag tttgatggca ccattagggt tagagatggt ggccatgggc gcatgtcctg      360

gccaactttg tatgatatat ggcagggtga ataggaaagt aaaattgtat tgtaaaaagg      420

gatttcttct gtttgttagc gcatgtacaa ggaatgcaag ttttgagcga gggggcatca      480

aagatctggc tgtgtttcca gctgtttttg ttagccccat cgaatccttg acataatgat      540

cccgcttaaa taagcaacct cgcttgtata gttccttgtg ctctaacaca cgatgatgat      600

aagtcgtaaa atagtggtgt ccaaagaatt tccaggccca gttgtaaaag ctaaaatgct      660

attcgaattt ctactagcag taagtcgtgt ttagaaatta tttttttata tacctttttt      720

ccttctatgt acagtaggac acagtgtcag cgccgcgttg acggagaata tttgcaaaaa      780

agtaaaagag aaagtcatag cggcgtatgt gccaaaaact tcgtcacaga gagggccata      840

agaaacatgg cccacggccc aatacgaagc accgcgacga agcccaaaca gcagtccgta      900

ggtggagcaa agcgctgggt aatacgcaaa cgttttgtcc caccttgact aatcacaaga      960

gtggagcgta ccttataaac cgagccgcaa gcaccgaatt gcgtacgcgt acgtgtggtt     1020

ttagagctat gctgttttgt ttttttt                                         1047


<210>  9
<211>  1087
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Maize optimized tracrRNA expression cassette

<400>  9
tgagagtaca atgatgaacc tagattaatc aatgccaaag tctgaaaaat gcaccctcag       60

tctatgatcc agaaaatcaa gattgcttga ggccctgttc ggttgttccg gattagagcc      120

ccggattaat tcctagccgg attacttctc taatttatat agattttgat gagctggaat      180

gaatcctggc ttattccggt acaaccgaac aggccctgaa ggataccagt aatcgctgag      240

ctaaattggc atgctgtcag agtgtcagta ttgcagcaag gtagtgagat aaccggcatc      300

atggtgccag tttgatggca ccattagggt tagagatggt ggccatgggc gcatgtcctg      360

gccaactttg tatgatatat ggcagggtga ataggaaagt aaaattgtat tgtaaaaagg      420

gatttcttct gtttgttagc gcatgtacaa ggaatgcaag ttttgagcga gggggcatca      480

aagatctggc tgtgtttcca gctgtttttg ttagccccat cgaatccttg acataatgat      540

cccgcttaaa taagcaacct cgcttgtata gttccttgtg ctctaacaca cgatgatgat      600

aagtcgtaaa atagtggtgt ccaaagaatt tccaggccca gttgtaaaag ctaaaatgct      660

attcgaattt ctactagcag taagtcgtgt ttagaaatta tttttttata tacctttttt      720

ccttctatgt acagtaggac acagtgtcag cgccgcgttg acggagaata tttgcaaaaa      780

agtaaaagag aaagtcatag cggcgtatgt gccaaaaact tcgtcacaga gagggccata      840

agaaacatgg cccacggccc aatacgaagc accgcgacga agcccaaaca gcagtccgta      900

ggtggagcaa agcgctgggt aatacgcaaa cgttttgtcc caccttgact aatcacaaga      960

gtggagcgta ccttataaac cgagccgcaa gcaccgaatt ggaaccattc aaaacagcat     1020

agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct     1080

ttttttt                                                               1087


<210>  10
<211>  39
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  crRNA containing a variable targeting domain targeting the 
       LIGCas-3 target sequence

<400>  10
gcguacgcgu acgugugguu uuagagcuau gcuguuuug                              39


<210>  11
<211>  86
<212>  RNA
<213>  Streptococcus pyogenes


<220>
<221>  misc_feature
<222>  (1)..(86)
<223>  tracrRNA

<400>  11
ggaaccauuc aaaacagcau agcaaguuaa aauaaggcua guccguuauc aacuugaaaa       60

aguggcaccg agucggugcu uuuuuu                                            86


<210>  12
<211>  24
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(24)
<223>  Maize genomic target site LIGCas-1 plus PAM sequence

<400>  12
gtaccgtacg tgccccggcg gagg                                              24


<210>  13
<211>  24
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(24)
<223>  Maize genomic target site LIGCas-2 plus PAM sequence

<400>  13
ggaattgtac cgtacgtgcc ccgg                                              24


<210>  14
<211>  20
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  Maize genomic target site LIGCas-3 plus PAM sequence

<400>  14
gcgtacgcgt acgtgtgagg                                                   20


<210>  15
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LIGCas-1 forward primer for primary PCR (crRNA-tracrRNA system)

<400>  15
ctacactctt tccctacacg acgctcttcc gatcttcctc tgtaacgatt tacgcacctg       60

ctg                                                                     63


<210>  16
<211>  58
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LIGCas-1 and LIGCas-2 reverse primer for primary PCR 
       (crRNA-tracrRNA system)

<400>  16
caagcagaag acggcatacg agctcttccg atctgcaaat gagtagcagc gcacgtat         58


<210>  17
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LIGCas-2 forward primer for primary PCR (crRNA-tracrRNA system)

<400>  17
ctacactctt tccctacacg acgctcttcc gatctgaagc tgtaacgatt tacgcacctg       60

ctg                                                                     63


<210>  18
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LIGCas-3 forward primer for primary PCR (crRNA-tracrRNA system)

<400>  18
ctacactctt tccctacacg acgctcttcc gatctaaggc gcaaatgagt agcagcgcac       60


<210>  19
<211>  57
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LIGCas-3 reverse primer for primary PCR (crRNA-tracrRNA and Long 
       guide RNA system)

<400>  19
caagcagaag acggcatacg agctcttccg atctcacctg ctgggaattg taccgta          57


<210>  20
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LIGCas-3 forward primer for primary PCR (Long guide RNA system)

<400>  20
ctacactctt tccctacacg acgctcttcc gatctttccc gcaaatgagt agcagcgcac       60


<210>  21
<211>  43
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Forward primer for secondary PCR

<400>  21
aatgatacgg cgaccaccga gatctacact ctttccctac acg                         43


<210>  22
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Reverse primer for secondary PCR

<400>  22
caagcagaag acggcata                                                     18


<210>  23
<211>  93
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(93)
<223>  Unmodified reference sequence for LIGCas-1 and LIGCas-2 locus

<400>  23
ctgtaacgat ttacgcacct gctgggaatt gtaccgtacg tgccccggcg gaggatatat       60

atacctcaca cgtacgcgta cgcgtatata tac                                    93


<210>  24
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 1 for LIGCas-1 locus

<400>  24
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggtcggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  25
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 2 for LIGCas-1

<400>  25
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggacggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  26
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 3 for LIGCas-1

<400>  26
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc gggcggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  27
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 4 for LIGCas-1

<400>  27
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggccggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  28
<211>  99
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(99)
<223>  Mutation 5 for LIGCas-1

<400>  28
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggatcggagg       60

atatatatac ctcacacgta cgcgtacgcg tatatatac                              99


<210>  29
<211>  94
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(94)
<223>  Mutation 6 for LIGCas-1

<400>  29
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggaggatata       60

tatacctcac acgtacgcgt acgcgtatat atac                                   94


<210>  30
<211>  81
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(81)
<223>  Mutation 7 for LIGCas-1

<400>  30
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggttcacacg       60

tacgcgtacg cgtatatata c                                                 81


<210>  31
<211>  65
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(65)
<223>  Mutation 8 for LIGCas-1

<400>  31
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtacgcg tacgcgtata       60

tatac                                                                   65


<210>  32
<211>  99
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(99)
<223>  Mutation 9 for LIGCas-1

<400>  32
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc ggttcggagg       60

atatatatac ctcacacgta cgcgtacgcg tatatatac                              99


<210>  33
<211>  95
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(95)
<223>  Mutation 10 for LIGCas-1

<400>  33
tcctctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcccc cggaggatat       60

atatacctca cacgtacgcg tacgcgtata tatac                                  95


<210>  34
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 1 for LIGCas-2

<400>  34
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgaccc cggcggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  35
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 2 for LIGCas-2

<400>  35
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgtccc cggcggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  36
<211>  96
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(96)
<223>  Mutation 3 for LIGCas-2

<400>  36
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacgtccccg gcggaggata       60

tatatacctc acacgtacgc gtacgcgtat atatac                                 96


<210>  37
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 4 for LIGCas-2

<400>  37
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacgtggccc cggcggagga       60

tatatatacc tcacacgtac gcgtacgcgt atatatac                               98


<210>  38
<211>  99
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 5 for LIGCas-2

<400>  38
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacgtgcacc ccggcggagg       60

atatatatac ctcacacgta cgcgtacgcg tatatatac                              99


<210>  39
<211>  87
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(87)
<223>  Mutation 6 for LIGCas-2

<400>  39
gaagctgtaa cgatttacgc acctgctggg aattgtaccc ggcggaggat atatatacct       60

cacacgtacg cgtacgcgta tatatac                                           87


<210>  40
<211>  92
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(92)
<223>  Mutation 7 for LIGCas-2

<400>  40
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tccccggcgg aggatatata       60

tacctcacac gtacgcgtac gcgtatatat ac                                     92


<210>  41
<211>  94
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(94)
<223>  Mutation 8 for LIGCas-2

<400>  41
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacccccggc ggaggatata       60

tatacctcac acgtacgcgt acgcgtatat atac                                   94


<210>  42
<211>  95
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(95)
<223>  Mutation 9 for LIGCas-2

<400>  42
gaagctgtaa cgatttacgc acctgctggg aattgtaccg tacgccccgg cggaggatat       60

atatacctca cacgtacgcg tacgcgtata tatac                                  95


<210>  43
<211>  88
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(88)
<223>  Mutation 10 for LIGCas-2

<400>  43
gaagctgtaa cgatttacgc acctgctggg aattgtaccc cggcggagga tatatatacc       60

tcacacgtac gcgtacgcgt atatatac                                          88


<210>  44
<211>  93
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(93)
<223>  Unmodified reference sequence for LIGCas-3 locus

<400>  44
cgcaaatgag tagcagcgca cgtatatata cgcgtacgcg tacgtgtgag gtatatatat       60

cctccgccgg ggcacgtacg gtacaattcc cag                                    93


<210>  45
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 1 for LIGCas-3 locus

<400>  45
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacgtt gtgaggtata       60

tatatcctcc gccggggcac gtacggtaca attcccag                               98


<210>  46
<211>  96
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(96)
<223>  Mutation 2 for LIGCas-3 locus

<400>  46
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacggt gaggtatata       60

tatcctccgc cggggcacgt acggtacaat tcccag                                 96


<210>  47
<211>  95
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(95)
<223>  Mutation 3 for LIGCas-3 locus

<400>  47
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacgtg aggtatatat       60

atcctccgcc ggggcacgta cggtacaatt cccag                                  95


<210>  48
<211>  96
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(96)
<223>  Mutation 4 for LIGCas-3 locus

<400>  48
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtactgt gaggtatata       60

tatcctccgc cggggcacgt acggtacaat tcccag                                 96


<210>  49
<211>  68
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(68)
<223>  Mutation 5 for LIGCas-3 locus

<400>  49
aaggcgcaaa tgagtagcag cgcacgtata tatatcctcc gccggggcac gtacggtaca       60

attcccag                                                                68


<210>  50
<211>  93
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(93)
<223>  Mutation 6 for LIGCas-3 locus

<400>  50
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtgtgag gtatatatat       60

cctccgccgg ggcacgtacg gtacaattcc cag                                    93


<210>  51
<211>  89
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(89)
<223>  Mutation 7 for LIGCas-3 locus

<400>  51
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgtgaggtat atatatcctc       60

cgccggggca cgtacggtac aattcccag                                         89


<210>  52
<211>  89
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(89)
<223>  Mutation 8 for LIGCas-3 locus

<400>  52
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtactat atatatcctc       60

cgccggggca cgtacggtac aattcccag                                         89


<210>  53
<211>  94
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(94)
<223>  Mutation 9 for LIGCas-3 locus

<400>  53
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacgga ggtatatata       60

tcctccgccg gggcacgtac ggtacaattc ccag                                   94


<210>  54
<211>  96
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(96)
<223>  Mutation 10 for LIGCas-3 locus

<400>  54
aaggcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacgat gaggtatata       60

tatcctccgc cggggcacgt acggtacaat tcccag                                 96


<210>  55
<211>  39
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(39)
<223>  crDNA sequence comprised of deoxyribonucleic acids (crDNA) 
       targeting

<400>  55
gcgtacgcgt acgtgtggtt ttagagctat gctgttttg                              39


<210>  56
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 1 for LIGCas-3 locus (synthetic crRNA plus tracrRNA and 
       Cas9

<400>  56
ggaacgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacgtt gtgaggtata       60

tatatcctcc gccggggcac gtacggtaca attcccag                               98


<210>  57
<211>  94
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(94)
<223>  Mutation 2 for LIGCas-3 locus (synthetic crRNA plus tracrRNA and 
       Cas9

<400>  57
ggaacgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtagtga ggtatatata       60

tcctccgccg gggcacgtac ggtacaattc ccag                                   94


<210>  58
<211>  96
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(96)
<223>  Mutation 3 for LIGCas-3 locus (synthetic crRNA plus tracrRNA and 
       Cas9

<400>  58
ggaacgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtactgt gaggtatata       60

tatcctccgc cggggcacgt acggtacaat tcccag                                 96


<210>  59
<211>  98
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(98)
<223>  Mutation 1 for LIGCas-3 locus (synthetic crDNA plus tracrRNA and 
       Cas9

<400>  59
ccttcgcaaa tgagtagcag cgcacgtata tatacgcgta cgcgtacgtt gtgaggtata       60

tatatcctcc gccggggcac gtacggtaca attcccag                               98


<210>  60
<211>  66
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(66)
<223>  Mutation 2 for LIGCas-3 locus (synthetic crDNA plus tracrRNA and 
       Cas9

<400>  60
ggaacgcaaa tgagtagcag cgcacgtata tatactccgc cggggcacgt acggtacaat       60

tcccag                                                                  66


<210>  61
<211>  85
<212>  DNA
<213>  Zea mays


<220>
<221>  misc_feature
<222>  (1)..(85)
<223>  Mutation 3 for LIGCas-3 locus (synthetic crDNA plus tracrRNA and 
       Cas9

<400>  61
ggaacgcaaa tgagtagcag cgcacgtata tatacgcgta cggtatatat atcctccgcc       60

ggggcacgta cggtacaatt cccag                                             85


<210>  62
<211>  17
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of unmodified crRNA VT domain

<400>  62
gcguacgcgu acgugug                                                      17


<210>  63
<211>  22
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of unmodified crRNA CER domain

<400>  63
guuuuagagc uaugcuguuu ug                                                22


<210>  64
<211>  17
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crRNA VT domain modified with Phosphorothioate bonds 
       at 5 prime end


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  The first N at the 5 prime end represents a G ribonucleotide with
       a phosphorothioate bond, the second  N represents a C 
       ribonucleotide with a phosphorothioate bond and third N 
       represents a G ribonucleotide with a Phosphorothioate bond.

<400>  64
nnnuacgcgu acgugug                                                      17


<210>  65
<211>  22
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crRNA CER domain modified with Phosphorothioate bonds
       near 3 prime end


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  the Ns at the nineteenth, twentieth and twenty-first position 
       represent U ribonucleotides with phosphorothioate bonds.

<400>  65
guuuuagagc uaugcugunn ng                                                22


<210>  66
<211>  17
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crRNA VT domain modified with 2-O-Methyl RNA bases at
       5 prime end


<220>
<221>  misc_feature
<223>  the first N at the 5 prime end represents a G 2-O-methyl 
       ribonucleotide, the second N represents a C 2-O-methyl 
       ribonucleotide and the third N represents a G 2-O-Methyl 
       ribonucleotide.

<220>
<221>  misc_feature
<222>  (1)..(3)
<223>  n is a, c, g, or u

<400>  66
nnnuacgcgu acgugug                                                      17


<210>  67
<211>  22
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crRNA CER domain modified with 2-O-Methyl RNA bases 
       at 3 prime end


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  the N at the twentieth position represents a U 2-O-Methyl 
       ribonucleotide, the N at the twenty-first position represents a U
       2-O-Methyl ribonucleotide and the N at the twenty-second position
       represents a G 2-O-Methyl ribonucleotide.

<400>  67
guuuuagagc uaugcuguun nn                                                22


<210>  68
<211>  17
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crRNA VT domain modified with 2-O-Methyl RNA bases 
       throughout


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  The first N at the 5 prime end represents a G 2-O-Methyl 
       ribonucleotide, a N at the second position represents a C 
       2-O-Methyl ribonucleotide, a N at the third position represents a
       G 2-O-Methyl ribonucleotide,

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the fourth position represents an U 2-O-Methyl 
       ribonucleotide, a N at the fifth position represents an A 
       2-O-Methyl ribonucleotide, a N at the sixth position represents a
       C 2-O-Methyl ribonucleotide,

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the seventh position represents a G 2-O-Methyl 
       ribonucleotide, a N at the eighth position represents a C 
       2'O-Methyl ribonucleotide, a N at the ninth position represents a
       G 2-O-Methyl ribonucleotide

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 
       position represents an U-, A-, C-, G-,U-, G-,U-, G- 2-O-Methyl 
       ribonucleotide, respectively.

<400>  68
nnnnnnnnnn nnnnnnn                                                      17


<210>  69
<211>  22
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crRNA CER domain modified with 2-O-Methyl RNA bases 
       throughout


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  The first N at the 5 prime end represents a G 2-O-Methyl 
       ribonucleotide, a N at the second position represents an U 
       2-O-Methyl ribonucleotide, a N at the third position represents 
       an U 2-O-Methyl ribonucleotide,

<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  a N at the 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 
       14th, 15th, 16th, 17th,18th, 19th, 20th, 21st, 22nd position 
       represents an U-, U-, A-, G-, A-, G-, C-, U-, A-, U-, G-, C-, U-,
       G, U-, U-, U-, U-, G-2-O-Methyl ribonucleotide, respectively.

<400>  69
nnnnnnnnnn nnnnnnnnnn nn                                                22


<210>  70
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of unmodified crDNA VT domain

<400>  70
gcgtacgcgt acgtgtg                                                      17


<210>  71
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of unmodified crDNA CER domain

<400>  71
gttttagagc tatgctgttt tg                                                22


<210>  72
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with one Locked Nucleic Acid
       base


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the sixteenth position represents a T Locked Nucleic Acid 
       base.

<400>  72
gcgtacgcgt acgtgng                                                      17


<210>  73
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with three Locked Nucleic 
       Acid 
bases


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a  N at the twelfth position represents a C Locked Nucleic Acid 
       base, a N at the fourteenth position represents a T Locked 
       Nucleic Acid base and a N at the sixteenth position represents a 
       T Locked Nucleic Acid base.

<400>  73
gcgtacgcgt angngng                                                      17


<210>  74
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with six Locked Nucleic Acid
       bases


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the 6th position represents a C Locked Nucleic Acid base, 
       a N at the 8th position represents a C Locked Nucleic Acid base, 
       a N at the tenth position represents a T Locked Nucleic Acid 
       base, a N at the twelfth position represents a C Locked Nucleic 
       

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the fourteenth position represents a T Locked Nucleic Acid
       base and a N at the sixteenth position represents a T Locked 
       Nucleic Acid base.

<400>  74
gcgtangngn angngng                                                      17


<210>  75
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with three Locked Nucleic 
       Acid bases and three phosphorothioate bonds


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a first N at the 5 prime end represents a G deoxyribonucleotide 
       with a phosphorothioate bond, a N at the second position  and a N
       at the third position represents a C-, and G-deoxyribonucleotide 
       with a phosphorothioate bond, respectively

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the twelfth position represents a C Locked Nucleic Acid 
       base, a N at the fourteenth position represents a T Locked 
       Nucleic Acid base and a N at the sixteenth position represents a 
       T Locked Nucleic Acid base.

<400>  75
nnntacgcgt angngng                                                      17


<210>  76
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA CER domain modified with three phosphorothioate
       bonds


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  the Ns at the nineteenth, twentieth and twenty-first positions 
       represent T deoxyribonucleotides with Phosphorothioate bonds.

<400>  76
gttttagagc tatgctgtnn ng                                                22


<210>  77
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with one 5-Methyl dC base


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a  N at the twelfth position represents a 5-Methyl dC 
       deoxyribonucleotide.

<400>  77
gcgtacgcgt angtgtg                                                      17


<210>  78
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with three 5-Methyl dC bases


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  Ns at the sixth, eighth and twelfth positions represent 5-Methyl 
       dC deoxyribonucleotides

<400>  78
gcgtangngt angtgtg                                                      17


<210>  79
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with one 2,6-Diaminopurine 
       base


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the eleventh position represents a 2,6-Diaminopurine 
       deoxyribonucleotide.

<400>  79
gcgtacgcgt ncgtgtg                                                      17


<210>  80
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain modified with two 2,6-Diaminopurine 
       base


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a Ns at the fifth and eleventh positions represent 
       2,6-diaminopurine deoxyribonucleotides

<400>  80
gcgtncgcgt ncgtgtg                                                      17


<210>  81
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain with Locked Nucleic Acid base 
       modifications at the 5 prime end


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  The first N at the 5 prime end represent a G Locked Nucleic Acid 
       base, second N represents a C Locked Nucleic Acid base and third 
       N represents a G Locked Nucleic Acid base.

<400>  81
nnntacgcgt acgtgtg                                                      17


<210>  82
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA CER domain with Locked Nucleic Acid base 
       modifications at the 3 prime end


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  A N at the twentieth position represents a T Locked Nucleic Acid 
       base, a N at the twenty-first position represents a T Locked 
       Nucleic Acid base and a N at the twenty-second position 
       represents a G Locked Nucleic Acid base.

<400>  82
gttttagagc tatgctgttn nn                                                22


<210>  83
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain with Phosphorothioate bond 
       modifications at the 5 prime end


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  The first N at the 5 prime end represents a G deoxyribonucleotide
       with a phosphorothioate bond, second N represents a C 
       deoxyribonucleotide with a phosphorothioate bond and third N 
       represents a G deoxyribonucleotide with a phosphorothioate bond.

<400>  83
nnntacgcgt acgtgtg                                                      17


<210>  84
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain with 2-O-Methyl RNA base 
       modifications at the 5 prime end


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  The first N at the 5 prime end represents a G 2-O-Methyl 
       ribonucleotide, second N represents a C 2-O-Methyl ribonucleotide
       and third N represents a G 2-O-Methyl ribonucleotide.

<400>  84
nnntacgcgt acgtgtg                                                      17


<210>  85
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA CER domain with 2-O-Methyl RNA base 
       modifications at the 3 prime end


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  a N at the twentieth position represents a U 2-O-Methyl 
       ribonucleotide, a N at the twenty-first position represents a U 
       2-O-Methyl ribonucleotide and a N at the twenty-second position 
       represent G 2-O-Methyl ribonucleotide.

<400>  85
gttttagagc tatgctgttn nn                                                22


<210>  86
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA VT domain with 2-O-Methyl RNA base 
       modifications at each position except T


<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  A first N at 5 prime end represents a G 2-O-Methyl 
       ribonucleotide, a N at the second position represents a C 
       2-O-Methyl ribonucleotide, a N at the third position represents a
       G 2-O-Methyl ribonucleotide,

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  a N at the  5th, 6th, 7th, 8th, 9th, 11th, 12th, 13th, 15th, 17th
       position represents an A-, C-, G-, C-, G-, A-, C-, G-, G-, G- 
       2'-O-Methyl ribonucleotide., respectively.

<400>  86
nnntnnnnnt nnntntn                                                      17


<210>  87
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of crDNA CER domain with 2-O-Methyl RNA base 
       modifications 
at each position except T


<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  a N at the sixth position represents an A 2-O-Methyl 
       ribonucleotide, a N at the seventh position represents a G 
       2-O-Methyl ribonucleotide, a N at the eighth position represents 
       an A 2-O-Methyl ribonucleotide,

<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  a N at the ninth position represents a G 2-O-Methyl 
       ribonucleotide, a N at the tenth position represents a C 
       2-O-Methyl ribonucleotide, a N at the twelfth position represents
       an A 2-O-Methyl ribonucleotide,

<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  a N at the fourteenth position represents a G 2-O-Methyl 
       ribonucleotide, a N at the fifteenth position represents a C 
       2-O-Methyl ribonucleotide, a N at the seventeenth position 
       represents a G 2-O-Methyl ribonucleotide

<220>
<221>  misc_feature
<222>  (1)..(22)
<223>  and a N at the twenty-second position represents a G 2-O-Methyl 
       ribonucleotide.

<400>  87
nttttnnnnn tntnntnttt tn                                                22


<210>  88
<211>  4104
<212>  DNA
<213>  artificial sequence

<220>
<223>  Saccharomyces cerevisiae codon optimized Cas9 coding sequence

<400>  88
atggataaga agtactccat tggcctagac atcggcacca attccgtggg ttgggccgtg       60

atcaccgacg agtacaaggt tccctccaag aagttcaagg tcttaggcaa taccgacagg      120

cactctatca agaagaatct gatcggtgct ttactgtttg actctggcga gaccgccgag      180

gccaccaggt tgaaacgtac cgctagaagg aggtacacca ggaggaagaa ccgtatctgc      240

tacctacaag agatcttctc caatgagatg gccaaggtcg acgactcctt cttccacagg      300

cttgaggagt ccttcctggt ggaggaggac aagaagcatg aaaggcaccc tatctttggc      360

aacatcgtcg acgaggttgc ctaccacgag aagtacccaa ccatctacca tctgaggaag      420

aaactggtag actccaccga caaggccgac ctgcgtctga tctacttagc cttagcccac      480

atgatcaagt ttagaggcca cttcctgatc gagggcgacc tgaatcccga taactccgac      540

gtcgataagc tgttcatcca gctggtccag acttacaatc agctgttcga ggagaacccc      600

atcaacgcct ctggcgtcga cgccaaagct atcctatcag cccgtctttc caagtctagg      660

aggctagaga acttgatcgc ccagcttccc ggcgagaaga agaacggcct gtttggcaat      720

ctgatcgccc tgtccttagg cttaactccc aacttcaaat ccaacttcga cctggccgaa      780

gatgctaagt tgcagttatc caaggacact tacgacgacg accttgataa cctgctggcc      840

caaatcggcg accagtacgc cgacctgttc ctagccgcca agaacttatc tgatgccatc      900

ttactgagtg acattctgag ggtcaacacc gagatcacca aagctccctt gagtgcctct      960

atgatcaaac gttacgacga acatcaccag gaccttaccc tgctaaaggc cctggtgagg     1020

caacaactgc ccgagaagta caaggagatc ttctttgacc agtccaagaa cggctacgcc     1080

ggctacatcg atggcggcgc ctctcaggag gagttctaca agttcatcaa gccaatctta     1140

gagaagatgg acggtaccga ggagctgctg gtaaagctga atagggagga cctgttaaga     1200

aaacagagga cctttgacaa cggctccatc cctcatcaga tccacctggg cgagctacac     1260

gccatcctgc gtcgtcagga ggacttctat cccttcctga aggacaacag agagaagatc     1320

gagaagattc tgacctttcg tataccctac tacgtgggcc ctctggccag aggcaattcc     1380

aggtttgcct ggatgaccag aaagtccgag gagaccatca caccctggaa ctttgaagag     1440

gttgtggaca agggcgcttc tgctcagtcc ttcatcgaga ggatgacaaa cttcgacaaa     1500

aacctgccca acgagaaggt cttacccaag cacagtctgc tgtacgagta cttcaccgta     1560

tacaacgagc tgaccaaggt taagtacgtc accgagggca tgagaaagcc tgccttcctg     1620

tccggagagc agaagaaggc cattgtggac ttgctgttca agaccaacag gaaagtgacc     1680

gtgaagcagc tgaaagagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

agtggcgtgg aggataggtt caacgcctcc ctaggcacat accacgatct actaaagatt     1800

atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctaga ggacatcgtc     1860

ttaaccttaa ccctattcga ggacagggag atgatcgagg agaggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaaacag ttgaagagga ggagatacac cggctggggc     1980

aggttatccc gtaagcttat caacggcatc cgtgacaagc agtccggcaa gaccatctta     2040

gacttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

tccctgactt tcaaggagga tatccagaag gctcaggtga gtggccaggg cgactcctta     2160

catgaacaca tcgcaaacct ggccggaagt cccgccatca agaagggcat cttgcaaact     2220

gtgaaagtgg ttgacgagct ggtgaaggtg atgggcaggc ataagcccga gaacatcgtt     2280

attgagatgg cccgtgagaa ccagacaacc cagaagggcc agaagaacag tcgtgaaagg     2340

atgaagagga tcgaggaagg catcaaggag ttaggctccc agatcctaaa ggagcacccc     2400

gtcgagaaca cccaactgca gaacgagaag ctgtacttgt actatctgca gaacggtagg     2460

gacatgtacg tcgaccagga gctggacatc aacaggctga gtgactacga cgtagaccac     2520

atcgtgccac agagtttcct gaaggacgac tctatcgaca acaaggtgct taccaggtcc     2580

gacaagaacc gtggcaagag tgacaatgtg ccctccgaag aggtggtcaa gaagatgaag     2640

aactactgga ggcagctgtt gaacgccaag ctgatcaccc agaggaagtt cgacaactta     2700

accaaagctg agagaggagg cctatctgaa ctagacaagg ccggtttcat caagaggcag     2760

ctggtcgaga ccaggcaaat caccaagcat gttgcccaga tcctagactc ccgtatgaac     2820

accaagtacg acgagaacga caagctaatc agagaggtga aggtgatcac cctgaagtcc     2880

aagctggtct ccgacttccg taaggacttc cagttctaca aggtcaggga gatcaacaac     2940

taccatcatg ctcatgacgc ctaccttaac gctgtggttg gcaccgcact gatcaagaag     3000

taccctaagt tggagagtga gttcgtatac ggcgactaca aggtctacga cgtgcgtaag     3060

atgatcgcca agtcagaaca ggaaatcggc aaggccaccg ccaagtactt cttctactct     3120

aacatcatga acttcttcaa aaccgagatc accctggcca acggcgagat ccgtaagagg     3180

cctcttatcg agaccaacgg cgaaaccggt gagatagtgt gggacaaggg cagagacttc     3240

gccaccgtga gaaaggtctt atccatgcct caagtcaaca ttgtcaagaa gaccgaggtc     3300

cagaccggcg gcttctctaa ggagagtatc ctgcccaagc gtaactctga caagctaatc     3360

gccagaaaga aggactggga ccctaagaag tatggaggct tcgactctcc cacagtagcc     3420

tactccgtgc tggtggtcgc caaggtcgag aagggcaaat ccaagaagct gaagtctgtg     3480

aaggagttgc taggcatcac catcatggag aggtcctcct tcgagaagaa tcccatcgac     3540

ttcttggagg ccaaaggtta caaggaggtc aagaaggacc tgatcatcaa gctgcccaag     3600

tactccttgt tcgagttaga gaacggcagg aagaggatgc tggcctccgc aggcgagtta     3660

cagaagggta acgagctggc cttaccctcc aagtacgtaa actttctgta cctggcttcc     3720

cattacgaga agctaaaggg cagtcccgag gacaacgagc agaagcagct gttcgtggag     3780

caacacaagc attacctgga cgagatcatc gagcagatct ccgagttcag taagcgtgtg     3840

atcctggccg acgccaatct ggacaaggtc ctgtccgcct acaacaagca tcgtgacaaa     3900

ccaatccgtg agcaggctga gaacatcatc catctattca ccctgaccaa cttaggcgcc     3960

cccgccgcct tcaagtactt tgataccact atcgacagga agaggtacac ctccaccaag     4020

gaagtactag acgccaccct gatccaccaa tccatcacag gcctgtacga gactaggatc     4080

gacttatccc agctaggtgg cgat                                            4104


<210>  89
<211>  20
<212>  DNA
<213>  Bacteriophage T7


<220>
<221>  T7 promoter sequence
<222>  (1)..(20)

<400>  89
taatacgact cactataggg                                                   20


<210>  90
<211>  20
<212>  DNA
<213>  Saccharomyces cerevisiae


<220>
<221>  ADE-URA3-DE2 target sequence minus PAM sequence
<222>  (1)..(20)

<400>  90
gcagacatta cgaatgcaca                                                   20


<210>  91
<211>  3387
<212>  DNA
<213>  S. thermophilus


<220>
<221>  misc_feature
<222>  (1)..(3387)
<223>  Cas9 endonuclease, genbank CS571758.1

<400>  91
atgagtgact tagttttagg acttgatatc ggtataggtt ctgttggtgt aggtatcctt       60

aacaaagtga caggagaaat tatccataaa aactcacgca tcttcccagc agctcaagca      120

gaaaataacc tagtacgtag aacgaatcgt caaggaagac gcttgacacg acgtaaaaaa      180

catcgtatag ttcgtttaaa tcgtctattt gaggaaagtg gattaatcac cgattttacg      240

aagatttcaa ttaatcttaa cccatatcaa ttacgagtta agggcttgac cgatgaattg      300

tctaatgaag aactgtttat cgctcttaaa aatatggtga aacaccgtgg gattagttac      360

ctcgatgatg ctagtgatga cggaaattca tcagtaggag actatgcaca aattgttaag      420

gaaaatagta aacaattaga aactaagaca ccgggacaga tacagttgga acgctaccaa      480

acatatggtc aattacgtgg tgattttact gttgagaaag atggcaaaaa acatcgcttg      540

attaatgtct ttccaacatc agcttatcgt tcagaagcct taaggatact gcaaactcaa      600

caagaattta atccacagat tacagatgaa tttattaatc gttatctcga aattttaact      660

ggaaaacgga aatattatca tggacccgga aatgaaaagt cacggactga ttatggtcgt      720

tacagaacga gtggagaaac tttagacaat atttttggaa ttctaattgg gaaatgtaca      780

ttttatccag aagagtttag agcagcaaaa gcttcctaca cggctcaaga attcaatttg      840

ctaaatgatt tgaacaatct aacagttcct actgaaacca aaaagttgag caaagaacag      900

aagaatcaaa tcattaatta tgtcaaaaat gaaaaggcaa tggggccagc gaaacttttt      960

aaatatatcg ctaagttact ttcttgtgat gttgcagata tcaagggata ccgtatcgac     1020

aaatcaggta aggctgagat tcatactttc gaagcctatc gaaaaatgaa aacgcttgaa     1080

accttagata ttgaacaaat ggatagagaa acgcttgata aattagccta tgtcttaaca     1140

ttaaacactg agagggaagg tattcaagaa gccttagaac atgaatttgc tgatggtagc     1200

tttagccaga agcaagttga cgaattggtt caattccgca aagcaaatag ttccattttt     1260

ggaaaaggat ggcataattt ttctgtcaaa ctgatgatgg agttaattcc agaattgtat     1320

gagacgtcag aagagcaaat gactatcctg acacgacttg gaaaacaaaa acgacttcgt     1380

cttcaaataa aacaaaatat ttcaaataaa acaaaatata tagatgagaa actattaact     1440

gaagaaatct ataatcctgt tgttgctaag tctgttcgcc aggctataaa aatcgtaaat     1500

gcggcgatta aagaatacgg agactttgac aatattgtca tcgaaatggc tcgtgaaaca     1560

aatgaagatg atgaaaagaa agctattcaa aagattcaaa aagccaacaa agatgaaaaa     1620

gatgcagcaa tgcttaaggc tgctaaccaa tataatggaa aggctgaatt accacatagt     1680

gttttccacg gtcataagca attagcgact aaaatccgcc tttggcatca gcaaggagaa     1740

cgttgccttt atactggtaa gacaatctca atccatgatt tgataaataa tcctaatcag     1800

tttgaagtag atcatatttt acctctttct atcacattcg atgatagcct tgcaaataag     1860

gttttggttt atgcaactgc taaccaagaa aaaggacaac gaacacctta tcaggcttta     1920

gatagtatgg atgatgcgtg gtctttccgt gaattaaaag cttttgtacg tgagtcaaaa     1980

acactttcaa acaagaaaaa agaatacctc cttacagaag aagatatttc aaagtttgat     2040

gttcgaaaga aatttattga acgaaatctt gtagatacaa gatacgcttc aagagttgtc     2100

ctcaatgccc ttcaagaaca ctttagagct cacaagattg atacaaaagt ttccgtggtt     2160

cgtggccaat ttacatctca attgagacgc cattggggaa ttgagaagac tcgtgatact     2220

tatcatcacc atgctgtcga tgcattgatt attgccgcct caagtcagtt gaatttgtgg     2280

aaaaaacaaa agaataccct tgtaagttat tcagaagaac aactccttga tattgaaaca     2340

ggtgaactta ttagtgatga tgagtacaag gaatctgtgt tcaaagcccc ttatcaacat     2400

tttgttgata cattgaagag taaagaattt gaagacagta tcttattctc atatcaagtg     2460

gattctaagt ttaatcgtaa aatatcagat gccactattt atgcgacaag acaggctaaa     2520

gtgggaaaag ataagaagga tgaaacttat gtcttaggga aaatcaaaga tatctatact     2580

caggatggtt atgatgcctt tatgaagatt tataagaagg ataagtcaaa attcctcatg     2640

tatcgtcacg acccacaaac ctttgagaaa gttatcgagc caattttaga gaactatcct     2700

aataagcaaa tgaatgaaaa aggaaaagag gtaccatgta atcctttcct aaaatataaa     2760

gaagaacatg gctatattcg taaatatagt aaaaaaggca atggtcctga aatcaagagt     2820

cttaaatact atgatagtaa gcttttaggt aatcctattg atattactcc agagaatagt     2880

aaaaataaag ttgtcttaca gtcattaaaa ccttggagaa cagatgtcta tttcaataag     2940

gctactggaa aatacgaaat ccttggatta aaatatgctg atctacaatt tgagaaaggg     3000

acaggaacat ataagatttc ccaggaaaaa tacaatgaca ttaagaaaaa agagggtgta     3060

gattctgatt cagaattcaa gtttacactt tataaaaatg atttgttact cgttaaagat     3120

acagaaacaa aagaacaaca gcttttccgt tttctttctc gaactttacc taaacaaaag     3180

cattatgttg aattaaaacc ttatgataaa cagaaatttg aaggaggtga ggcgttaatt     3240

aaagtgttgg gtaacgttgc taatggtggt caatgcataa aaggactagc aaaatcaaat     3300

atttctattt ataaagtaag aacagatgtc ctaggaaatc agcatatcat caaaaatgag     3360

ggtgataagc ctaagctaga tttttaa                                         3387


<210>  92
<211>  3369
<212>  DNA
<213>  S. thermophilus


<220>
<221>  misc_feature
<222>  (1)..(3369)
<223>  Cas9 endonuclease, genbank CS571770.1

<400>  92
atgagtgact tagttttagg acttgatatc ggtataggtt ctgttggtgt aggtatcctt       60

aacaaagtga caggagaaat tatccataaa aactcacgca tcttcccagc agctcaagca      120

gaaaataacc tagtacgtag aacgaatcgt caaggaagac gcttgacacg acgtaaaaaa      180

catcgtatag ttcgtttaaa tcgtctattt gaggaaagtg gattaatcac cgattttacg      240

aagatttcaa ttaatcttaa cccatatcaa ttacgagtta agggcttgac cgatgaattg      300

tctaatgaag aactgtttat cgctcttaaa aatatggtga aacaccgtgg gattagttac      360

ctcgatgatg ctagtgatga cggaaattca tcagtaggag actatgcaca aattgttaag      420

gaaaatagta aacaattaga aactaagaca ccgggacaga tacagttgga acgctaccaa      480

acatatggtc aattacgtgg tgattttact gttgagaaag atggcaaaaa acatcgcttg      540

attaatgtct ttccaacatc agcttatcgt tcagaagcct taaggatact gcaaactcaa      600

caagaattta attcacagat tacagatgaa tttattaatc gttatctcga aattttaact      660

ggaaaacgga aatattatca tggacccgga aatgaaaagt cacggactga ttatggtcgt      720

tacagaacga atggagaaac tttagacaat atttttggaa ttctaattgg gaaatgtaca      780

ttttatccag acgagtttag agcagcaaaa gcttcctaca cggctcaaga attcaatttg      840

ctaaatgatt tgaacaatct aacagttcct actgaaacca aaaagttgag caaagaacag      900

aagaatcaaa tcattaatta tgtcaaaaat gaaaaggtaa tggggccagc gaaacttttt      960

aaatatatcg ctaaattact ttcttgtgat gttgcagata tcaagggaca ccgtatcgac     1020

aaatcaggta aggctgagat tcatactttc gaagcctatc gaaaaatgaa aacgcttgaa     1080

accttagata ttgagcaaat ggatagagaa acgcttgata aattagccta tgtcttaaca     1140

ttaaacactg agagggaagg tattcaagaa gctttagaac atgaatttgc tgatggtagc     1200

tttagccaga agcaagttga cgaattggtt caattccgca aagcaaatag ttccattttt     1260

ggaaaaggat ggcataattt ttctgtcaaa ctgatgatgg agttaattcc agaattgtat     1320

gagacgtcag aagagcaaat gactatcctg acacgacttg gaaaacaaaa aacaacttcg     1380

tcttcaaata aaacaaaata tatagatgag aaactattaa ctgaagaaat ctataatcct     1440

gttgttgcta agtctgttcg ccaggctata aaaatcgtaa atgcggcgat taaagaatac     1500

ggagactttg acaatattgt catcgaaatg gctcgtgaaa caaatgaaga tgatgaaaag     1560

aaagctattc aaaagattca aaaagccaac aaagatgaaa aagatgcagc aatgcttaag     1620

gctgctaacc aatataatgg aaaggctgaa ttaccacata gtgttttcca cggtcataag     1680

caattagcga ctaaaatccg cctttggcat cagcaaggag aacgttgcct ttatactggt     1740

aagacaatct caatccatga tttgataaat aatcctaatc agtttgaagt agatcatatt     1800

ttacctcttt ctatcacatt cgatgatagc cttgcaaata aggttttggt ttatgcaact     1860

gctaaccaag aaaaaggaca acgaacacct tatcaggctt tagatagtat ggatgatgcg     1920

tggtctttcc gtgaattaaa agcttttgta cgtgagtcaa aaacactttc aaacaagaaa     1980

aaagaatacc tccttacaga agaagatatt tcaaagtttg atgttcgaaa gaaatttatt     2040

gaacgaaatc ttgtagatac aagatacgct tcaagagttg tcctcaatgc ccttcaagaa     2100

cactttagag ctcacaagat tgatacaaaa gtttccgtgg ttcgtggcca atttacatct     2160

caattgagac gccattgggg aattgagaag actcgtgata cttatcatca ccatgctgtc     2220

gatgcattga ttattgccgc ctcaagtcag ttgaatttgt ggaaaaaaca aaagaatacc     2280

cttgtaagtt attcagaaga acaactcctt gatattgaaa caggtgaact tattagtgat     2340

gatgagtaca aggaatctgt gttcaaagcc ccttatcaac attttgttga tacattgaag     2400

agtaaagaat ttgaagacag tatcttattc tcatatcaag tggattctaa gtttaatcgt     2460

aaaatatcag atgccactat ttatgcgaca agacaggcta aagtgggaaa agataagaag     2520

gatgaaactt atgtcttagg gaaaatcaaa gatatctata ctcaggatgg ttatgatgcc     2580

tttatgaaga tttataagaa ggataagtca aaattcctca tgtatcgtca cgacccacaa     2640

acctttgaga aagttatcga gccaatttta gagaactatc ctaataagga aatgaatgaa     2700

aaagggaaag aagtaccatg taatcctttc ctaaaatata aagaagaaca tggctatatt     2760

cgtaaatata gtaaaaaagg caatggtcct gaaatcaaga gtcttaaata ctatgatagt     2820

aagcttttag gtaatcctat tgatattact ccagagaata gtaaaaataa agttgtctta     2880

cagtcattaa aaccttggag aacagatgtc tatttcaata aaaatactgg taaatatgaa     2940

attttaggac tgaaatatgc tgatttacaa tttgaaaaga agacaggaac atataagatt     3000

tcccaggaaa aatacaatgg cattatgaaa gaagagggtg tagattctga ttcagaattc     3060

aagtttacac tttataaaaa tgatttgtta ctcgttaaag atacagaaac aaaagaacaa     3120

cagcttttcc gttttctttc tcgaactatg cctaatgtga aatattatgt agagttaaag     3180

ccttattcaa aagataaatt tgagaagaat gagtcactta ttgaaatttt aggttctgca     3240

gataagtcag gacgatgtat aaaagggcta ggaaaatcaa atatttctat ttataaggta     3300

agaacagatg tcctaggaaa tcagcatatc atcaaaaatg agggtgataa gcctaagcta     3360

gatttttaa                                                             3369


<210>  93
<211>  4113
<212>  DNA
<213>  S. agalactiae


<220>
<221>  misc_feature
<222>  (1)..(4113)
<223>  Cas9 endonuclease, genbank CS571785.1

<400>  93
atgaataagc catattcaat aggccttgac atcggtacta attccgtcgg atggagcatt       60

attacagatg attataaagt acctgctaag aagatgagag ttttagggaa cactgataaa      120

gaatatatta agaagaatct cataggtgct ctgctttttg atggcgggaa tactgctgca      180

gatagacgct tgaagcgaac tgctcgtcgt cgttatacac gtcgtagaaa tcgtattcta      240

tatttacaag aaatttttgc agaggaaatg agtaaagttg atgatagttt ctttcatcga      300

ttagaggatt cttttctagt tgaggaagat aagagaggga gcaagtatcc tatctttgca      360

acattgcagg aagagaaaga ttatcatgaa aaattttcga caatctatca tttgagaaaa      420

gaattagctg acaagaaaga aaaagcagac cttcgtctta tttatattgc tctagctcat      480

atcattaaat ttagagggca tttcctaatt gaggatgata gctttgatgt caggaataca      540

gacatttcaa aacaatatca agatttttta gaaatcttta atacaacttt tgaaaataat      600

gatttgttat ctcaaaacgt tgacgtagag gcaatactaa cagataagat tagcaagtct      660

gcgaagaaag atcgtatttt agcgcagtat cctaaccaaa aatctactgg catttttgca      720

gaatttttga aattgattgt cggaaatcaa gctgacttca agaaatattt caatttggag      780

gataaaacgc cgcttcaatt cgctaaggat agctacgatg aagatttaga aaatcttctt      840

ggacagattg gtgatgaatt tgcagactta ttctcagcag cgaaaaagtt atatgatagt      900

gtccttttgt ctggcattct tacagtaatc gacctcagta ccaaggcgcc actttcagct      960

tctatgattc agcgttatga tgaacataga gaggacttga aacagttaaa acaattcgta     1020

aaagcttcat tgccggaaaa atatcaagaa atatttgctg attcatcaaa agatggctac     1080

gctggttata ttgaaggtaa aactaatcaa gaagcttttt ataaatacct gtcaaaattg     1140

ttgaccaagc aagaagatag cgagaatttt cttgaaaaaa tcaagaatga agatttcttg     1200

agaaaacaaa ggacctttga taatggctca attccacacc aagtccattt gacagagctg     1260

aaagctatta tccgccgtca atcagaatac tatcccttct tgaaagagaa tcaagatagg     1320

attgaaaaaa tccttacctt tagaattcct tattatatcg ggccactagc acgtgagaag     1380

agtgattttg catggatgac tcgcaaaaca gatgacagta ttcgaccttg gaattttgaa     1440

gacttggttg ataaagaaaa atctgcggaa gcttttatcc atcgtatgac caacaatgat     1500

ttttatcttc ctgaagaaaa agttttacca aagcatagtc ttatttatga aaaatttacg     1560

gtctataatg agttgactaa ggttagatat aaaaatgagc aaggtgagac ttattttttt     1620

gatagcaata ttaaacaaga aatctttgat ggagtattca aggaacatcg taaggtatcc     1680

aagaagaagt tgctagattt tctggctaaa gaatatgagg agtttaggat agtagatgtt     1740

attggtctag ataaagaaaa taaagctttc aacgcctcat tgggaactta ccacgatctc     1800

gaaaaaatac tagacaaaga ttttctagat aatccagata atgagtctat tctggaagat     1860

atcgtccaaa ctctaacatt atttgaagac agagaaatga ttaagaagcg tcttgaaaac     1920

tataaagatc tttttacaga gtcacaacta aaaaaactct atcgtcgtca ctatactggc     1980

tggggacgat tgtctgctaa gttaatcaat ggtattcgag ataaagagag tcaaaaaaca     2040

atcttggact atcttattga tgatggtaga tctaatcgca actttatgca gttgataaat     2100

gatgatggtc tatctttcaa atcaattatc agtaaggcac aggctggtag tcattcagat     2160

aatctaaaag aagttgtagg tgagcttgca ggtagccctg ctattaaaaa gggaattcta     2220

caaagtttga aaattgttga tgagcttgtt aaagtcatgg gatacgaacc tgaacaaatt     2280

gtggttgaga tggcgcgtga gaatcaaaca acaaatcaag gtcgtcgtaa ctctcgacaa     2340

cgctataaac ttcttgatga tggcgttaag aatctagcta gtgacttgaa tggcaatatt     2400

ttgaaagaat atcctacgga taatcaagcg ttgcaaaatg aaagactttt cctttactac     2460

ttacaaaacg gaagagatat gtatacaggg gaagctctag atattgacaa tttaagtcaa     2520

tatgatattg accacattat tcctcaagct ttcataaaag atgattctat tgataatcgt     2580

gttttggtat catctgctaa aaatcgtgga aagtcagatg atgttcctag ccttgaaatt     2640

gtaaaagatt gtaaagtttt ctggaaaaaa ttacttgatg ctaagttaat gagtcagcgt     2700

aagtatgata atttgactaa ggcagagcgc ggaggcctaa cttccgatga taaggcaaga     2760

tttatccaac gtcagttggt tgagacacga caaattacca agcatgttgc ccgtatcttg     2820

gatgaacgct ttaataatga gcttgatagt aaaggtagaa ggatccgcaa agttaaaatt     2880

gtaaccttga agtcaaattt ggtttcaaat ttccgaaaag aatttggatt ctataaaatt     2940

cgtgaagtta acaattatca ccatgcacat gatgcctatc ttaatgcagt agttgctaaa     3000

gctattctaa ccaaatatcc tcagttagag ccagaatttg tctacggcga ctatccaaaa     3060

tataatagtt acaaaacgcg taaatccgct acagaaaagc tatttttcta ttcaaatatt     3120

atgaacttct ttaaaactaa ggtaacttta gcggatggaa ccgttgttgt aaaagatgat     3180

attgaagtta ataatgatac gggtgaaatt gtttgggata aaaagaaaca ctttgcgaca     3240

gttagaaaag tcttgtcata ccctcagaac aatatcgtga agaagacaga gattcagaca     3300

ggtggtttct ctaaggaatc aatcttggcg catggtaact cagataagtt gattccaaga     3360

aaaacgaagg atatttattt agatcctaag aaatatggag gttttgatag tccgatagta     3420

gcttactctg ttttagttgt agctgatatc aaaaagggta aagcacaaaa actaaaaaca     3480

gttacggaac ttttaggaat taccatcatg gagaggtcca gatttgagaa aaatccatca     3540

gctttccttg aatcaaaagg ctatttaaat attagggctg ataaactaat tattttgccc     3600

aagtatagtc tgttcgaatt agaaaatggg cgtcgtcgat tacttgctag tgctggtgaa     3660

ttacaaaaag gtaatgagct agccttacca acacaattta tgaagttctt ataccttgca     3720

agtcgttata atgagtcaaa aggtaaacca gaggagattg agaagaaaca agaatttgta     3780

aatcaacatg tctcttattt tgatgacatc cttcaattaa ttaatgattt ttcaaaacga     3840

gttattctag cagatgctaa tttagagaaa atcaataagc tttaccaaga taataaggaa     3900

aatatatcag tagatgaact tgctaataat attatcaatc tatttacttt taccagtcta     3960

ggagctccag cagcttttaa attttttgat aaaatagttg atagaaaacg ctatacatca     4020

actaaagaag tacttaattc taccctaatt catcaatcta ttactggact ttatgaaaca     4080

cgtattgatt tgggtaagtt aggagaagat tga                                  4113


<210>  94
<211>  4134
<212>  DNA
<213>  S. agalactiae


<220>
<221>  misc_feature
<222>  (1)..(4134)
<223>  Cas9 endonuclease, genbank CS571790.1

<400>  94
atgaataagc catattcaat aggccttgac atcggtacta attccgtcgg atggagcatt       60

attacagatg attataaagt acctgctaag aagatgagag ttttagggaa cactgataaa      120

gaatatatta agaagaatct cataggtgct ctgctttttg atggcgggaa tactgctgca      180

gatagacgct tgaagcgaac tgctcgtcgt cgttatacac gtcgtagaaa tcgtattcta      240

tatttacaag aaatttttgc agaggaaatg agtaaagttg atgatagttt ctttcatcga      300

ttagaggatt cttttctagt tgaggaagat aagagaggta gcaagtatcc tatctttgca      360

acaatgcagg aggagaaata ttatcatgaa aaatttccga caatctatca tttgagaaaa      420

gaattggctg acaagaaaga aaaagcagac cttcgtcttg tttatctggc tctagctcat      480

atcattaaat tcagagggca tttcctaatt gaggatgata gatttgatgt gaggaatacc      540

gatattcaaa aacaatatca agccttttta gaaatttttg atactacctt tgaaaataat      600

catttgttat ctcaaaatgt agatgtagaa gcaattctaa cagataagat tagcaagtct      660

gcgaagaagg atcgcatctt agcgcagtat cctaaccaaa aatctactgg tatttttgca      720

gaatttttga aattgattgt cggaaatcaa gctgacttca agaaacattt caatttggag      780

gataaaacac cgcttcaatt cgctaaggat agctacgatg aagatttaga aaatcttctt      840

ggacagattg gtgatgaatt tgcagactta ttctcagtag cgaaaaagct atatgatagt      900

gttcttttat ctggcattct tacagtaact gatctcagta ccaaggcgcc actttctgcc      960

tctatgattc agcgttatga tgaacatcat gaggacttaa agcatctaaa acaattcgta     1020

aaagcttcat tacctgaaaa ttatcgggaa gtatttgctg attcatcaaa agatggctac     1080

gctggctata ttgaaggcaa aactaatcaa gaagcttttt ataaatatct gttaaaattg     1140

ttgaccaaac aagaaggtag cgagtatttt cttgagaaaa ttaagaatga agattttttg     1200

agaaaacaga gaacctttga taatggctca atcccgcatc aagtccattt gacagaattg     1260

agggctatta ttcgacgtca atcagaatac tatccattct tgaaagagaa tcaagatagg     1320

attgaaaaaa tccttacctt tagaattcct tattatgtcg ggccactagc acgtgagaag     1380

agtgattttg catggatgac tcgcaaaaca gatgacagta ttcgaccttg gaattttgaa     1440

gacttggttg ataaagaaaa atctgcggaa gcttttatcc atcgcatgac caacaatgac     1500

ctctatcttc cagaagaaaa agttttacca aagcatagtc ttatttatga aaaatttact     1560

gtttacaatg aattaacgaa ggttagattt ttggcagaag gctttaaaga ttttcaattt     1620

ttaaatagga agcaaaaaga aactatcttt aacagcttgt ttaaggaaaa acgtaaagta     1680

actgaaaagg atattattag ttttttgaat aaagttgatg gatatgaagg aattgcaatc     1740

aaaggaattg agaaacagtt taacgctagc ctttcaacct atcatgatct taaaaaaata     1800

cttggcaagg atttccttga taatacagat aacgagctta ttttggaaga tatcgtccaa     1860

actctaacct tatttgaaga tagagaaatg attaagaagt gtcttgacat ctataaagat     1920

ttttttacag agtcacagct taaaaagctc tatcgccgtc actatactgg ctggggacga     1980

ttgtctgcta agctaataaa tggcatccga aataaagaga atcaaaaaac aatcttggac     2040

tatcttattg atgatggaag tgcaaaccga aacttcatgc agttgataaa tgatgatgat     2100

ctatcattta aaccaattat tgacaaggca cgaactggta gtcattcgga taatctgaaa     2160

gaagttgtag gtgaacttgc tggtagccct gctattaaaa aagggattct acaaagtttg     2220

aaaatagttg atgagctggt taaagtcatg ggctatgaac ctgaacaaat cgtggttgaa     2280

atggcacgtg agaaccaaac gacagcaaaa ggattaagtc gttcacgaca acgcttgaca     2340

accttgagag aatctcttgc taatttgaag agtaatattt tggaagagaa aaagcctaag     2400

tatgtgaaag atcaagttga aaatcatcat ttatctgatg accgtctttt cctttactac     2460

ttacaaaacg gaagagatat gtatacaaaa aaggctctgg atattgataa tttaagtcaa     2520

tatgatattg accacattat tcctcaagct ttcataaaag atgattctat tgataatcgt     2580

gttttggtat catctgctaa aaatcgtgga aaatcagatg atgttcctag cattgaaatt     2640

gtaaaagctc gcaaaatgtt ctggaaaaat ttactggatg ctaagttaat gagtcagcgt     2700

aagtatgata atttgactaa ggcagagcgc ggaggcctaa cttccgatga taaggcaaga     2760

tttatccaac gtcagttggt tgagactcga caaattacca agcatgtagc tcgtatcttg     2820

gatgaacgct tcaataatga agttgataat ggtaaaaaga tttgcaaggt taaaattgta     2880

accttgaagt caaatttggt ttcaaatttc cgaaaagaat ttggattcta taaaattcgt     2940

gaagttaatg attatcacca tgcacacgat gcttatctta atgcagtagt tgccaaagct     3000

attctaacca aatatccaca gttagagcca gagtttgtct acggaatgta tagacagaaa     3060

aaactttcga aaatcgttca tgaggataag gaagaaaaat atagtgaagc aaccaggaaa     3120

atgtttttct actccaactt gatgaatatg ttcaaaagag ttgtgaggtt agcagatggt     3180

tctattgttg taagaccagt aatagaaact ggtagatata tgagaaaaac tgcatgggat     3240

aaaaagaaac actttgcgac agttagaaaa gtcttgtcat accctcagaa caatatcgtg     3300

aagaagacag agattcagac aggtggtttc tctaaggaat caatcttggc gcatggtaac     3360

tcagataagt tgattccaag aaaaacgaag gatatttatt tagatcctaa gaaatatgga     3420

ggttttgata gtccgatagt agcttactct gttttagttg tagctgatat caaaaaaggt     3480

aaagcacaaa aactaaaaac agttacggaa cttttaggaa ttaccatcat ggagaggtcc     3540

agatttgaga aaaatccatc agctttcctt gaatcaaaag gttatttaaa tattagggac     3600

gataaattaa tgattttacc gaagtatagt ctgttcgaat tagaaaatgg gcgtcgtcga     3660

ttacttgcta gtgctggtga attacaaaaa ggtaacgagc tagccttacc aacacaattt     3720

atgaagttct tataccttgc aagtcgttat aatgagtcaa aaggtaaacc agaggagatt     3780

gagaagaaac aagaatttgt aaatcaacat gtctcttatt ttgatgacat ccttcaatta     3840

attaatgatt tttcaaaacg agttattcta gcagatgcta atttagagaa aatcaataag     3900

ctttaccagg ataataagga aaatatacca gtagatgaac ttgctaataa tattatcaat     3960

ctatttactt ttaccagtct aggagctcca gcagctttta aattttttga taaaatagtt     4020

gatagaaaac gctatacatc aactaaagaa gtacttaatt ctactctaat ccatcaatct     4080

attactggac tttatgaaac acgtattgat ttgggtaaat taggagaaga ttga           4134


<210>  95
<211>  4038
<212>  DNA
<213>  S. mutans


<220>
<221>  misc_feature
<222>  (1)..(4038)
<223>  Cas9 endonuclease, genbank CS571790.1

<400>  95
atgaaaaaac cttactctat tggacttgat attggaacca attctgttgg ttgggctgtt       60

gtgacagatg actacaaagt tcctgctaag aagatgaagg ttctgggaaa tacagataaa      120

agtcatatcg agaaaaattt gcttggcgct ttattatttg atagcgggaa tactgcagaa      180

gacagacggt taaagagaac tgctcgccgt cgttacacac gtcgcagaaa tcgtatttta      240

tatttgcaag agattttttc agaagaaatg ggcaaggtag atgatagttt ctttcatcgt      300

ttagaggatt cttttcttgt tactgaggat aaacgaggag agcgccatcc catttttggg      360

aatcttgaag aagaagttaa gtatcatgaa aattttccaa ccatttatca tttgcggcaa      420

tatcttgcgg ataatccaga aaaagttgat ttgcgtttag tttatttggc tttggcacat      480

ataattaagt ttagaggtca ttttttaatt gaaggaaagt ttgatacacg caataatgat      540

gtacaaagac tgtttcaaga atttttagca gtctatgata atacttttga gaatagttcg      600

cttcaggagc aaaatgttca agttgaagaa attctgactg ataaaatcag taaatctgct      660

aagaaagata gagttttgaa actttttcct aatgaaaagt ctaatggccg ctttgcagaa      720

tttctaaaac taattgttgg taatcaagct gattttaaaa agcattttga attagaagag      780

aaagcaccat tgcaattttc taaagatact tatgaagaag agttagaagt actattagct      840

caaattggag ataattacgc agagctcttt ttatcagcaa agaaactgta tgatagtatc      900

cttttatcag ggattttaac agttactgat gttggtacca aagcgccttt atctgcttcg      960

atgattcagc gatataatga acatcagatg gatttagctc agcttaaaca attcattcgt     1020

cagaaattat cagataaata taacgaagtt ttttctgatg tttcaaaaga cggctatgcg     1080

ggttatattg atgggaaaac aaatcaagaa gctttttata aataccttaa aggtctatta     1140

aataagattg agggaagtgg ctatttcctt gataaaattg agcgtgaaga ttttctaaga     1200

aagcaacgta cctttgacaa tggctctatt ccacatcaga ttcatcttca agaaatgcgt     1260

gctatcattc gtagacaggc tgaattttat ccgtttttag cagacaatca agataggatt     1320

gagaaattat tgactttccg tattccctac tatgttggtc cattagcgcg cggaaaaagt     1380

gattttgctt ggttaagtcg gaaatcggct gataaaatta caccatggaa ttttgatgaa     1440

atcgttgata aagaatcctc tgcagaagct tttatcaatc gtatgacaaa ttatgatttg     1500

tacttgccaa atcaaaaagt tcttcctaaa catagtttat tatacgaaaa atttactgtt     1560

tacaatgaat taacaaaggt taaatataaa acagagcaag gaaaaacagc attttttgat     1620

gccaatatga agcaagaaat ctttgatggc gtatttaagg tttatcgaaa agtaactaaa     1680

gataaattaa tggatttcct tgaaaaagaa tttgatgaat ttcgtattgt tgatttaaca     1740

ggtctggata aagaaaataa agtatttaac gcttcttatg gaacttatca tgatttgtgt     1800

aaaattttag ataaagattt tctcgataat tcaaagaatg aaaagatttt agaagatatt     1860

gtgttgacct taacgttatt tgaagataga gaaatgatta gaaaacgtct agaaaattac     1920

agtgatttat tgaccaaaga acaagtgaaa aagctggaaa gacgtcatta tactggttgg     1980

ggaagattat cagctgagtt aattcatggt attcgcaata aagaaagcag aaaaacaatt     2040

cttgattatc tcattgatga tggcaatagc aatcggaact ttatgcaact gattaacgat     2100

gatgctcttt ctttcaaaga agagattgct aaggcacaag ttattggaga aacagacaat     2160

ctaaatcaag ttgttagtga tattgctggc agccctgcta ttaaaaaagg aattttacaa     2220

agcttgaaga ttgttgatga gcttgtcaaa attatgggac atcaacctga aaatatcgtc     2280

gtggagatgg cgcgtgaaaa ccagtttacc aatcagggac gacgaaattc acagcaacgt     2340

ttgaaaggtt tgacagattc tattaaagaa tttggaagtc aaattcttaa agaacatccg     2400

gttgagaatt cacagttaca aaatgataga ttgtttctat attatttaca aaacggcaga     2460

gatatgtata ctggagaaga attggatatt gattatctaa gccagtatga tatagaccat     2520

attatcccgc aagcttttat aaaggataat tctattgata atagagtatt gactagctca     2580

aaggaaaatc gtggaaaatc ggatgatgta ccaagtaaag atgttgttcg taaaatgaaa     2640

tcctattgga gtaagctact ttcggcaaag cttattacac aacgtaaatt tgataatttg     2700

acaaaagctg aacgaggtgg attgaccgac gatgataaag ctggattcat caagcgtcaa     2760

ttagtagaaa cacgacaaat taccaaacat gtagcacgta ttctggacga acgatttaat     2820

acagaaacag atgaaaacaa caagaaaatt cgtcaagtaa aaattgtgac cttgaaatca     2880

aatcttgttt ccaatttccg taaagagttt gaactctaca aagtgcgtga aattaatgac     2940

tatcatcatg cacatgatgc ctatctcaat gctgtaattg gaaaggcttt actaggtgtt     3000

tacccacaat tggaacctga atttgtttat ggtgattatc ctcattttca tggacataaa     3060

gaaaataaag caactgctaa gaaatttttc tattcaaata ttatgaactt ctttaaaaaa     3120

gatgatgtcc gtactgataa aaatggtgaa attatctgga aaaaagatga gcatatttct     3180

aatattaaaa aagtgctttc ttatccacaa gttaatattg ttaagaaagt agaggagcaa     3240

acgggaggat tttctaaaga atctatcttg ccgaaaggta attctgacaa gcttattcct     3300

cgaaaaacga agaaatttta ttgggatacc aagaaatatg gaggatttga tagcccgatt     3360

gttgcttatt ctattttagt tattgctgat attgaaaaag gtaaatctaa aaaattgaaa     3420

acagtcaaag ccttagttgg tgtcactatt atggaaaaga tgacttttga aagggatcca     3480

gttgcttttc ttgagcgaaa aggctatcga aatgttcaag aagaaaatat tataaagtta     3540

ccaaaatata gtttatttaa actagaaaac ggacgaaaaa ggctattggc aagtgctagg     3600

gaacttcaaa agggaaatga aatcgttttg ccaaatcatt taggaacctt gctttatcac     3660

gctaaaaata ttcataaagt tgatgaacca aagcatttgg actatgttga taaacataaa     3720

gatgaattta aggagttgct agatgttgtg tcaaactttt ctaaaaaata tactttagca     3780

gaaggaaatt tagaaaaaat caaagaatta tatgcacaaa ataatggtga agatcttaaa     3840

gaattagcaa gttcatttat caacttatta acatttactg ctataggagc accggctact     3900

tttaaattct ttgataaaaa tattgatcga aaacgatata cttcaactac tgaaattctc     3960

aacgctaccc tcatccacca atccatcacc ggtctttatg aaacgcggat tgatctcaat     4020

aagttaggag gagactaa                                                   4038


<210>  96
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  single guide RNA targeting the LIGCas-3 target sequence

<400>  96
gcguacgcgu acgugugguu uuagagcuag aaauagcaag uuaaaauaag gcuaguccgu       60

uaucaacuug aaaaaguggc accgagucgg ugcu                                   94


