                         SEQUENCE LISTING

<110>  DSM IP Assets B.V.
 
<120>  CRISPR Transient Expression Construct (CTEC)

<130>  32805-WO-PCT

<150>  18171496.5
<151>  2018-05-09

<150>  18184210.5
<151>  2018-07-18

<160>  171   

<170>  PatentIn version 3.5

<210>  1
<211>  5441
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Cas9, including a C-terminal SV40 nuclear 
       localization signal, codon pair optimized for expression in 
       Saccharomyces cerevisiae

<400>  1
ttttcttttt ttgcggtcac ccccatgtgg cggggaggca gaggagtagg tagagcaacg       60

aatcctacta tttatccaaa ttagtctagg aactcttttt ctagattttt tagatttgag      120

ggcaagcgct gttaacgact cagaaatgta agcactacgg agtagaacga gaaatccgcc      180

ataggtggaa atcctagcaa aatcttgctt accctagcta gcctcaggta agctagcctt      240

agcctgtcaa atttttttca aaatttggta agtttctact agcaaagcaa acacggttca      300

acaaaccgaa aactccactc attatacgtg gaaaccgaaa caaaaaaaca aaaaccaaaa      360

tactcgccaa tgagaaagtt gctgcgtttc tactttcgag gaagaggaac tgagaggatt      420

gactacgaaa ggggcaaaaa cgagtcgtat tctcccatta ttgtctgcta ccacgcggtc      480

tagtagaata agcaaccagt caacgctaag acaggtaatc aaaataccag tctgctggct      540

acgggctagt ttttacctct tttagaaccc actgtaaaag tccgttgtaa agcccgttct      600

cactgttggc gttttttttt ttttggttta gtttcttatt tttcattttt ttctttcatg      660

accaaaaaca aacaaatctc gcgatttgta ctgcggccac tggggcgtgg ccaaaaaaat      720

gacaaattta gaaaccttag tttctgattt ttcctgttat gaggagatat gataaaaaat      780

attactgctt tattgttttt tttttatcta ctgaaataga gaaacttacc caaggaggag      840

gcaaaaaaaa gagtatatat acagcagcta ccattcagat tttaatatat tcttttctct      900

tcttctacac tattattata ataattttac tatattcatt tttagcttaa aacctcatag      960

aatattattc ttcagtcact cgcttaaata cttatcaaaa atggacaaga aatactctat     1020

tggtttggat atcgggacca actccgtcgg ttgggctgtc atcaccgacg aatacaaggt     1080

tccatccaag aaattcaagg tcttgggtaa cactgacaga cactctatca agaagaattt     1140

gatcggtgct ttgttgttcg actccggtga aaccgctgaa gctaccagat tgaagcgtac     1200

cgctcgtcgt agatacacta gacgtaaaaa ccgtatttgt tacttgcaag aaatcttttc     1260

taacgaaatg gccaaggttg acgactcttt cttccacaga ttggaagaat ctttcttggt     1320

tgaagaagac aagaagcacg aaagacatcc aatcttcggt aacatcgttg acgaagttgc     1380

ttaccacgaa aaatacccta ccatctacca tttgagaaag aagttggtcg attccaccga     1440

caaggctgat ttgagattga tctatttggc cttggctcac atgatcaagt tcagaggtca     1500

cttcttgatt gaaggtgact tgaacccaga caactctgac gtcgacaaat tgttcatcca     1560

attggtccaa acctacaacc aattattcga ggaaaaccca attaacgctt ctggtgttga     1620

tgctaaggcc atcttatctg cccgtttgtc caagtctaga cgtttggaaa acttgattgc     1680

tcaattgcct ggtgaaaaga aaaacggttt gttcggtaac ttgatcgctt tgtccttggg     1740

tttgacccca aacttcaagt ccaacttcga cttggctgaa gatgccaagt tgcaattgtc     1800

caaggacacc tacgacgacg acttagacaa cttgttggct caaatcggtg accaatacgc     1860

cgacttgttc ttggctgcca aaaacttatc tgacgctatc ttgttgtctg acatcttgag     1920

agttaacact gaaattacca aggctccatt gtctgcttct atgatcaaaa gatacgacga     1980

acaccaccaa gatctgactt tgttgaaggc tttggttaga caacaattgc cagaaaagta     2040

caaggaaatc ttcttcgacc aatccaaaaa tggttacgcc ggttacattg acggtggtgc     2100

ttctcaggaa gaattctaca agttcatcaa gccaattttg gaaaagatgg atggtactga     2160

agaattattg gttaagttga acagagaaga cttattgaga aagcaacgta ccttcgataa     2220

cggttctatc ccacaccaaa tccacttggg tgaattgcac gccattttga gaagacagga     2280

agatttctat ccattcctaa aggacaacag agaaaagatc gaaaagatct taactttcag     2340

aatcccatac tacgtcggtc cattggccag aggtaattct agattcgctt ggatgaccag     2400

aaagtctgaa gaaaccatca ccccatggaa cttcgaagaa gtcgtcgaca agggtgcttc     2460

tgcccaatct ttcatcgaaa gaatgaccaa ctttgataag aacttgccaa acgagaaggt     2520

cttgccaaag cactctttgt tgtacgaata cttcaccgtc tacaacgaat taaccaaggt     2580

taaatacgtt actgaaggta tgagaaagcc agctttccta tccggtgaac aaaagaaggc     2640

tattgttgac ttgttgttta agaccaacag aaaggtcact gttaagcaat tgaaggaaga     2700

ctacttcaag aagattgaat gtttcgattc cgtcgaaatc tccggtgttg aagaccgttt     2760

caatgcttct ttgggcacct accacgattt gttaaagatc atcaaggaca aggacttttt     2820

agataacgaa gaaaacgaag acatcttgga agatatcgtt ttgaccttga ctcttttcga     2880

ggacagagaa atgattgaag agagattgaa gacctacgct cacttgttcg acgataaagt     2940

tatgaagcaa ctaaagagaa gaagatacac tggttggggt agattgtcca gaaagttgat     3000

taacggtatc agagacaagc aatccggtaa gactatttta gactttttga aatccgatgg     3060

tttcgctaac agaaacttta tgcaattgat tcacgacgat tctttgactt tcaaggaaga     3120

cattcaaaaa gcccaagtct ctggtcaagg tgattctttg cacgaacaca tcgctaactt     3180

ggctggttct ccagctatta agaagggtat cttacaaacc gtcaaggtcg ttgatgaatt     3240

ggtcaaagtc atgggtagac acaagccaga aaatattgtc atcgaaatgg ctagagaaaa     3300

ccaaactact caaaagggtc aaaagaactc tagagaacgt atgaagagaa ttgaagaagg     3360

tatcaaggag ttgggttctc aaattttgaa agaacaccca gtcgaaaaca ctcaattaca     3420

aaacgaaaag ctatacttgt actacttgca aaacggtcgt gacatgtacg tcgaccaaga     3480

attggatatc aacagattgt ctgactacga tgtcgatcat atcgtcccac aatcgttctt     3540

gaaggacgat tccattgaca acaaagtttt gactagatct gacaagaaca gaggtaagtc     3600

tgataacgtt ccatctgaag aagttgttaa gaagatgaag aactactgga gacaattgtt     3660

gaatgctaag ttgatcactc aaagaaagtt cgacaacttg accaaggctg aaagaggtgg     3720

tttgtccgaa ttggacaaag ccggtttcat caagagacaa ttagtcgaaa ctagacaaat     3780

caccaagcat gttgctcaaa tcttggattc cagaatgaac actaagtacg atgaaaacga     3840

caaactaatt agagaagtta aggtcatcac tttgaagtct aagttggttt ctgacttcag     3900

aaaggacttc caattttaca aggtcagaga aatcaacaac taccatcacg ctcacgatgc     3960

ctacttgaac gctgttgtcg gtactgcctt aatcaaaaag tacccaaagt tggaatctga     4020

attcgtttac ggtgactaca aggtttacga tgttagaaag atgatcgcca agtctgaaca     4080

agaaattggt aaggccactg ctaagtactt cttctactct aacatcatga actttttcaa     4140

gactgaaatc actttagcta acggtgaaat tagaaagcgt ccattgattg aaaccaatgg     4200

tgaaactggt gaaattgtct gggacaaggg tagagatttc gctaccgtca gaaaggtttt     4260

gtctatgcca caagttaaca tcgtcaagaa gactgaagtt caaactggtg gtttctctaa     4320

ggaatccatt ttgccaaaga gaaactctga caagttgatt gctagaaaga aggactggga     4380

tcctaagaag tacggtggtt tcgactctcc aactgttgct tactccgttt tggtcgttgc     4440

taaggttgaa aagggtaagt ctaagaagtt gaagtctgtt aaggaattgt tgggtatcac     4500

catcatggaa agatcctcct tcgaaaagaa cccaatcgac tttttggaag ctaagggtta     4560

caaggaagtc aagaaggatt tgatcattaa gttaccaaaa tactccttgt tcgaattgga     4620

aaacggtaga aagagaatgt tggcctccgc tggtgaacta caaaaaggta acgaattggc     4680

tttaccatct aagtacgtta acttcttgta cttggcttcc cactacgaaa agttgaaagg     4740

ttccccagaa gacaacgaac aaaagcaatt gtttgttgaa caacacaagc actacttgga     4800

tgaaattatt gaacaaatct ccgaattctc caagagagtc attttggctg atgctaactt     4860

agataaggtt ttatccgctt acaacaagca cagagacaaa ccaatcagag aacaagctga     4920

aaacatcatt catttgttca ctttaaccaa cttgggtgct ccagctgctt tcaaatactt     4980

cgacactacc attgacagaa agagatacac ttccaccaaa gaagttttag atgctacttt     5040

gattcaccaa tctattaccg gtttgtacga aaccagaatt gacttgtctc aattgggtgg     5100

tgattccaga gctgatccaa agaagaagag aaaggtgtaa aggagttaaa ggcaaagttt     5160

tcttttctag agccgttccc acaaataatt atacgtatat gcttcttttc gtttactata     5220

tatctatatt tacaagcctt tattcactga tgcaatttgt ttccaaatac ttttttggag     5280

atctcataac tagatatcat gatggcgcaa cttggcgcta tcttaattac tctggctgcc     5340

aggcccgtgt agagggccgc aagaccttct gtacgccata tagtctctaa gaacttgaac     5400

aagtttctag acctattgcc gcctttcgga tcgctattgt t                         5441


<210>  2
<211>  11742
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of vector pCSN061

<400>  2
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>  3
<211>  5712
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of vector pRN1120

<400>  3
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>  4
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to obtain 
       Pthd3-YFP-TenoI expression cassette

<400>  4
gtgcttagtc aaaaaattag ccttttaatt c                                      31


<210>  5
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to obtain 
       Pthd3-YFP-TenoI expression cassette

<400>  5
gaggggagga aatgagaaat gag                                               23


<210>  6
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to attach connector 5 
       to the Pthd3-YFP-TenoI expression cassette

<400>  6
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt gtgcttagtc       60

aaaaaattag cc                                                           72


<210>  7
<211>  73
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to attach connector 3 
       to the Pthd3-YFP-TenoI expression cassette

<400>  7
acttagtatg gtctgttgga aaggattgtg gcttcgcata caggctttct gaggggagga       60

aatgagaaat gag                                                          73


<210>  8
<211>  1730
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the Pthd3-YFP-TenoI expression cassette 
       flanked by connector 5 (CON5) and connector 3 (CON3); 
       CON5-Pthd3-YFP-TenoI-CON3

<400>  8
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 cccctccctc     1680

agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt                1730


<210>  9
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to attach a 50 bp 
       genomic DNA flank to connector 5 of YFP expression cassette; 
       CON5-Pthd3-YFP-TenoI-CON3

<400>  9
cttcatgcca gcaatagttg cgtgctgagc tcaacagtgc ccaacccttg aagcgacttc       60

caatcgcttt gc                                                           72


<210>  10
<211>  74
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to attach a 50 bp 
       genomic DNA flank to connector 3 of YFP expression cassette; 
       CON5-Pthd3-YFP-TenoI-CON3

<400>  10
gaaaagcact cctttagtac cactcaacaa gttgtctgat gacaaagaat acttagtatg       60

gtctgttgga aagg                                                         74


<210>  11
<211>  1830
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CON5-Pthd3-YFP-TenoI-CON3 expression 
       cassette that contains 50 bp genomic DNA flanks at 5' and 3' side
       for integration in the genome

<400>  11
cttcatgcca gcaatagttg cgtgctgagc tcaacagtgc ccaacccttg aagcgacttc       60

caatcgcttt gcatatccag taccacaccc acaggcgttt gtgcttagtc aaaaaattag      120

ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta cacagaatat      180

ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa tataatggag      240

cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa tattgttttc      300

ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga acaggggcac      360

aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg gagtaaatga      420

tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc ttctattacc      480

ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt tccctgaaat      540

tattccccta cttgactaat aagtatataa agacggtagg tattgattgt aattctgtaa      600

atctatttct taaacttctt aaattctact tttatagtta gtcttttttt tagttttaaa      660

acaccaagaa cttagtttcg aataaacaca cataaacaaa caaaatgtct aaaggtgaag      720

aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca      780

aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat      840

tgatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttaggtt      900

atggtttgca atgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt      960

ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact     1020

acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa     1080

aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata     1140

actctcacaa tgtttacatc actgctgaca aacaaaagaa tggtatcaaa gctaacttca     1200

aaattagaca caacattgaa gatggtggtg ttcaattagc tgaccattat caacaaaata     1260

ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatctg     1320

ccttatccaa agatccaaac gaaaagagag atcacatggt cttgttagaa tttgttactg     1380

ctgctggtat tacccatggt atggatgaat tgtacaaata aaagcttttg attaagcctt     1440

ctagtccaaa aaacacgttt ttttgtcatt tatttcattt tcttagaata gtttagttta     1500

ttcattttat agtcacgaat gttttatgat tctatatagg gttgcaaaca agcatttttc     1560

attttatgtt aaaacaattt caggtttacc ttttattctg cttgtggtga cgcgtgtatc     1620

cgcccgctct tttggtcacc catgtattta attgcataaa taattcttaa aagtggagct     1680

agtctatttc tatttacata cctctcattt ctcatttcct cccctccctc agaaagcctg     1740

tatgcgaagc cacaatcctt tccaacagac catactaagt attctttgtc atcagacaac     1800

ttgttgagtg gtactaaagg agtgcttttc                                      1830


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the guide sequence (genomic target 
       sequence) of INT1 for Cas9

<400>  12
tattagaacc agggaggtcc                                                   20


<210>  13
<211>  488
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the complete guide RNA cassette for 
       targeting CAS9 to INT1 locus in the genome that contains homology
       to vector backbone pRN1120 for homologous recombination

<400>  13
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct attagaacca gggaggtccg ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtctgg ggggcccggt acccagcttt tgttcccttt agtgagggtt      480

aattccga                                                               488


<210>  14
<211>  488
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to INT1 and donor DNA on the 3' side

<400>  14
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct attagaacca gggaggtccg ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtctac aaatctgcaa ccccagcttc ataagctttc      420

tctcccacca gcaaagcatg gacctccctg gttctaataa tgagcgactg aagttttcca      480

aaagaaac                                                               488


<210>  15
<211>  538
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-2 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to INT1, connector A and donor DNA on 
       the 3' side

<400>  15
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct attagaacca gggaggtccg ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtcttt gcccatcgaa cgtacaagta ctcctctgtt      420

ctctccttcc tttgctttac aaatctgcaa ccccagcttc ataagctttc tctcccacca      480

gcaaagcatg gacctccctg gttctaataa tgagcgactg aagttttcca aaagaaac        538


<210>  16
<211>  488
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-3 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to INT1 and donor DNA on the 5' side

<400>  16
acaaatctgc aaccccagct tcataagctt tctctcccac cagcaaagca tggacctccc       60

tggttctaat aatgagcgac tgaagttttc caaaagaaac tctttgaaaa gataatgtat      120

gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag tatatacaag      180

gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct tgggctagcg      240

gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt tggtcaaacg      300

ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg cagtgaaaga      360

taaatgatct attagaacca gggaggtccg ttttagagct agaaatagca agttaaaata      420

aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt      480

ttatgtct                                                               488


<210>  17
<211>  538
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-4 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to INT1, connector A and donor DNA on 
       the 5' side

<400>  17
acaaatctgc aaccccagct tcataagctt tctctcccac cagcaaagca tggacctccc       60

tggttctaat aatgagcgac tgaagttttc caaaagaaac ttgcccatcg aacgtacaag      120

tactcctctg ttctctcctt cctttgcttt tctttgaaaa gataatgtat gattatgctt      180

tcactcatat ttatacagaa acttgatgtt ttctttcgag tatatacaag gtgattacat      240

gtacgtttga agtacaactc tagattttgt agtgccctct tgggctagcg gtaaaggtgc      300

gcattttttc acaccctaca atgttctgtt caaaagattt tggtcaaacg ctgtagaagt      360

gaaagttggt gcgcatgttt cggcgttcga aacttctccg cagtgaaaga taaatgatct      420

attagaacca gggaggtccg ttttagagct agaaatagca agttaaaata aggctagtcc      480

gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt ttatgtct        538


<210>  18
<211>  511
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-5 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to INT1, PAM and guide target sequence
       and donor DNA on the 5' side

<400>  18
acaaatctgc aaccccagct tcataagctt tctctcccac cagcaaagca tggacctccc       60

tggttctaat aatgagcgac tgaagttttc caaaagaaac cctggacctc cctggttcta      120

atatctttga aaagataatg tatgattatg ctttcactca tatttataca gaaacttgat      180

gttttctttc gagtatatac aaggtgatta catgtacgtt tgaagtacaa ctctagattt      240

tgtagtgccc tcttgggcta gcggtaaagg tgcgcatttt ttcacaccct acaatgttct      300

gttcaaaaga ttttggtcaa acgctgtaga agtgaaagtt ggtgcgcatg tttcggcgtt      360

cgaaacttct ccgcagtgaa agataaatga tctattagaa ccagggaggt ccgttttaga      420

gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga      480

gtcggtggtg ctttttttgt tttttatgtc t                                     511


<210>  19
<211>  511
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-6B comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to INT1, PAM and guide target sequence
       and donor DNA on the 3' side

<400>  19
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct attagaacca gggaggtccg ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtctcc tggacctccc tggttctaat aacaaatctg      420

caaccccagc ttcataagct ttctctccca ccagcaaagc atggacctcc ctggttctaa      480

taatgagcga ctgaagtttt ccaaaagaaa c                                     511


<210>  20
<211>  499
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA)  for Cas9 targeting to the YFP gene and donor DNA on the 
       3' side

<400>  20
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtctat ttgtactact ggtaaattgc cagttccatg      420

gccaacctta gtcactactt tagttatggt ttgcaatgtt ttgctagata cccagatcat      480

atgaaacaac atgactttt                                                   499


<210>  21
<211>  549
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-2 comprising a guide RNA cassette 
       (sgRNA)  for Cas9 targeting to the YFP gene, connector A and 
       donor DNA on the 3' side

<400>  21
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtcttt gcccatcgaa cgtacaagta ctcctctgtt      420

ctctccttcc tttgctttat ttgtactact ggtaaattgc cagttccatg gccaacctta      480

gtcactactt tagttatggt ttgcaatgtt ttgctagata cccagatcat atgaaacaac      540

atgactttt                                                              549


<210>  22
<211>  499
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-3 comprising a guide RNA 
       cassette(sgRNA)  for Cas9 targeting to the YFP gene and donor DNA
       on the 5' side

<400>  22
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttagttatg       60

gtttgcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt ttctttgaaa      120

agataatgta tgattatgct ttcactcata tttatacaga aacttgatgt tttctttcga      180

gtatatacaa ggtgattaca tgtacgtttg aagtacaact ctagattttg tagtgccctc      240

ttgggctagc ggtaaaggtg cgcatttttt cacaccctac aatgttctgt tcaaaagatt      300

ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt tcggcgttcg aaacttctcc      360

gcagtgaaag ataaatgatc ttagtcacta ctttaggtta gttttagagc tagaaatagc      420

aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct      480

ttttttgttt tttatgtct                                                   499


<210>  23
<211>  549
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-4 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to the YFP gene, connector A and donor
       DNA on the 5' side

<400>  23
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttagttatg       60

gtttgcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt tttgcccatc      120

gaacgtacaa gtactcctct gttctctcct tcctttgctt ttctttgaaa agataatgta      180

tgattatgct ttcactcata tttatacaga aacttgatgt tttctttcga gtatatacaa      240

ggtgattaca tgtacgtttg aagtacaact ctagattttg tagtgccctc ttgggctagc      300

ggtaaaggtg cgcatttttt cacaccctac aatgttctgt tcaaaagatt ttggtcaaac      360

gctgtagaag tgaaagttgg tgcgcatgtt tcggcgttcg aaacttctcc gcagtgaaag      420

ataaatgatc ttagtcacta ctttaggtta gttttagagc tagaaatagc aagttaaaat      480

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct ttttttgttt      540

tttatgtct                                                              549


<210>  24
<211>  522
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nnucleotide sequence of CTEC-5 comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to the YFP gene, PAM and guide target 
       sequence and donor DNA on the 5' side

<400>  24
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttagttatg       60

gtttgcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt tccataacct      120

aaagtagtga ctaatctttg aaaagataat gtatgattat gctttcactc atatttatac      180

agaaacttga tgttttcttt cgagtatata caaggtgatt acatgtacgt ttgaagtaca      240

actctagatt ttgtagtgcc ctcttgggct agcggtaaag gtgcgcattt tttcacaccc      300

tacaatgttc tgttcaaaag attttggtca aacgctgtag aagtgaaagt tggtgcgcat      360

gtttcggcgt tcgaaacttc tccgcagtga aagataaatg atcttagtca ctactttagg      420

ttagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa      480

agtggcaccg agtcggtggt gctttttttg ttttttatgt ct                         522


<210>  25
<211>  522
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-6A comprising a guide RNA cassette 
       (sgRNA) for Cas9 targeting to the YFP gene, guide target and PAM 
       sequence and donor DNA on the 3' side

<400>  25
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtcttt agtcactact ttaggttatg gatttgtact      420

actggtaaat tgccagttcc atggccaacc ttagtcacta ctttagttat ggtttgcaat      480

gttttgctag atacccagat catatgaaac aacatgactt tt                         522


<210>  26
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of guide sequence (genomic target sequence) 
       of INT1 for Cas9

<400>  26
tattagaacc agggaggtcc                                                   20


<210>  27
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of guide sequence (genomic target sequence) 
       of YFP for Cas9

<400>  27
ttagtcacta ctttaggtta                                                   20


<210>  28
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of connector A

<400>  28
ttgcccatcg aacgtacaag tactcctctg ttctctcctt cctttgcttt                  50


<210>  29
<211>  388
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the complete guide RNA expression cassette
       for targeting Cas9 to the YFP expression cassette in the genome 
       of CSN009

<400>  29
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtct                                         388


<210>  30
<211>  388
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the complete guide RNA expression cassette
       for targeting Cas9 to the INT1 locus in the genome of CSN001

<400>  30
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct attagaacca gggaggtccg ttttagagct      300

agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc      360

ggtggtgctt tttttgtttt ttatgtct                                         388


<210>  31
<211>  111
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the YFP donor DNA that is part of CTEC 
       fragments for Cas9 editing

<400>  31
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttagttatg       60

gtttgcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt t               111


<210>  32
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the INT1 donor DNA that is part of CTEC 
       fragments for Cas9 editing

<400>  32
acaaatctgc aaccccagct tcataagctt tctctcccac cagcaaagca tggacctccc       60

tggttctaat aatgagcgac tgaagttttc caaaagaaac                            100


<210>  33
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify CTEC 
       fragments that contain donor DNA on the 3' side

<400>  33
tctttgaaaa gataatgtat gattatgc                                          28


<210>  34
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify CTEC 
       fragments that contain the YFP donor DNA on the 5' side

<400>  34
atttgtacta ctggtaaatt gccag                                             25


<210>  35
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments that contain the YFP donor DNA on the 3' side

<400>  35
aaaagtcatg ttgtttcata tgatctg                                           27


<210>  36
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments that contain donor DNA on the 5' side

<400>  36
agacataaaa aacaaaaaaa gcaccacc                                          28


<210>  37
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify CTEC 
       fragments that contain the INT1 donor DNA on the 5' side

<400>  37
acaaatctgc aaccccagct tc                                                22


<210>  38
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments that contain the INT1 donor DNA on the 3' side

<400>  38
gtttcttttg gaaaacttca gtcgctc                                           27


<210>  39
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify the YFP ORF

<400>  39
atgtctaaag gtgaagaatt attcac                                            26


<210>  40
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify the YFP ORF

<400>  40
ttttatttgt acaattcatc catacc                                            26


<210>  41
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of forward primer used for sequencing the YFP
       ORF

<400>  41
ttttatttgt acaattcatc catacc                                            26


<210>  42
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify part of the 
       INT1 locus

<400>  42
attaagtaat agatacgcac aacc                                              24


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

<220>
<223>  Nucleotide sequence of the reverse primer to amplify part of the 
       INT1 locus

<400>  43
ggaatactac cagatcgatc c                                                 21


<210>  44
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer used for sequencing 
       part of the INT1 locus

<400>  44
attaagtaat agatacgcac aacc                                              24


<210>  45
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify the 
       Kl11p-pCSN061 backbone-GND2t PCR fragment

<400>  45
ttttgataag tatttaagcg agtg                                              24


<210>  46
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify the 
       Kl11p-pCSN061 backbone-GND2t PCR fragment

<400>  46
aggagttaaa ggcaaagttt tc                                                22


<210>  47
<211>  1239
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Protein sequence of LbCpf1 (from Lachnospiraceae bacterium 
       ND2006) including a C-terminal NLS

<400>  47

Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr 
1               5                   10                  15      


Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp 
            20                  25                  30          


Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys 
        35                  40                  45              


Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp 
    50                  55                  60                  


Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu 
65                  70                  75                  80  


Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn 
                85                  90                  95      


Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn 
            100                 105                 110         


Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu 
        115                 120                 125             


Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe 
    130                 135                 140                 


Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn 
145                 150                 155                 160 


Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile 
                165                 170                 175     


Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys 
            180                 185                 190         


Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys 
        195                 200                 205             


Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe 
    210                 215                 220                 


Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile 
225                 230                 235                 240 


Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn 
                245                 250                 255     


Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys 
            260                 265                 270         


Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser 
        275                 280                 285             


Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe 
    290                 295                 300                 


Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys 
305                 310                 315                 320 


Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile 
                325                 330                 335     


Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe 
            340                 345                 350         


Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp 
        355                 360                 365             


Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp 
    370                 375                 380                 


Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu 
385                 390                 395                 400 


Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu 
                405                 410                 415     


Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser 
            420                 425                 430         


Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys 
        435                 440                 445             


Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys 
    450                 455                 460                 


Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr 
465                 470                 475                 480 


Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile 
                485                 490                 495     


Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr 
            500                 505                 510         


Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro 
        515                 520                 525             


Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala 
    530                 535                 540                 


Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys 
545                 550                 555                 560 


Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly 
                565                 570                 575     


Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met 
            580                 585                 590         


Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro 
        595                 600                 605             


Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly 
    610                 615                 620                 


Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys 
625                 630                 635                 640 


Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn 
                645                 650                 655     


Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu 
            660                 665                 670         


Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys 
        675                 680                 685             


Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile 
    690                 695                 700                 


Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His 
705                 710                 715                 720 


Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile 
                725                 730                 735     


Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys 
            740                 745                 750         


Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys 
        755                 760                 765             


Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr 
    770                 775                 780                 


Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile 
785                 790                 795                 800 


Ala Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val 
                805                 810                 815     


Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Asp 
            820                 825                 830         


Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly 
        835                 840                 845             


Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn 
    850                 855                 860                 


Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu 
865                 870                 875                 880 


Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile 
                885                 890                 895     


Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 
            900                 905                 910         


Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu Asn 
        915                 920                 925             


Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr Gln 
    930                 935                 940                 


Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp Lys 
945                 950                 955                 960 


Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln Ile 
                965                 970                 975     


Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly Phe 
            980                 985                 990         


Ile Phe Tyr Ile Pro Ala Trp Leu  Thr Ser Lys Ile Asp  Pro Ser Thr 
        995                 1000                 1005             


Gly Phe  Val Asn Leu Leu Lys  Thr Lys Tyr Thr Ser  Ile Ala Asp 
    1010                 1015                 1020             


Ser Lys  Lys Phe Ile Ser Ser  Phe Asp Arg Ile Met  Tyr Val Pro 
    1025                 1030                 1035             


Glu Glu  Asp Leu Phe Glu Phe  Ala Leu Asp Tyr Lys  Asn Phe Ser 
    1040                 1045                 1050             


Arg Thr  Asp Ala Asp Tyr Ile  Lys Lys Trp Lys Leu  Tyr Ser Tyr 
    1055                 1060                 1065             


Gly Asn  Arg Ile Arg Ile Phe  Arg Asn Pro Lys Lys  Asn Asn Val 
    1070                 1075                 1080             


Phe Asp  Trp Glu Glu Val Cys  Leu Thr Ser Ala Tyr  Lys Glu Leu 
    1085                 1090                 1095             


Phe Asn  Lys Tyr Gly Ile Asn  Tyr Gln Gln Gly Asp  Ile Arg Ala 
    1100                 1105                 1110             


Leu Leu  Cys Glu Gln Ser Asp  Lys Ala Phe Tyr Ser  Ser Phe Met 
    1115                 1120                 1125             


Ala Leu  Met Ser Leu Met Leu  Gln Met Arg Asn Ser  Ile Thr Gly 
    1130                 1135                 1140             


Arg Thr  Asp Val Asp Phe Leu  Ile Ser Pro Val Lys  Asn Ser Asp 
    1145                 1150                 1155             


Gly Ile  Phe Tyr Asp Ser Arg  Asn Tyr Glu Ala Gln  Glu Asn Ala 
    1160                 1165                 1170             


Ile Leu  Pro Lys Asn Ala Asp  Ala Asn Gly Ala Tyr  Asn Ile Ala 
    1175                 1180                 1185             


Arg Lys  Val Leu Trp Ala Ile  Gly Gln Phe Lys Lys  Ala Glu Asp 
    1190                 1195                 1200             


Glu Lys  Leu Asp Lys Val Lys  Ile Ala Ile Ser Asn  Lys Glu Trp 
    1205                 1210                 1215             


Leu Glu  Tyr Ala Gln Thr Ser  Val Lys His Ser Arg  Ala Asp Pro 
    1220                 1225                 1230             


Lys Lys  Lys Arg Lys Val 
    1235                 


<210>  48
<211>  3720
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence CPO LbCpf1 including a C-terminal NLS

<400>  48
atgtctaagt tggaaaaatt caccaactgt tactctttgt ctaagacttt gagattcaag       60

gccatcccag ttggtaagac ccaagaaaac atcgacaaca agagactatt agttgaagat      120

gaaaagagag ctgaagacta caagggtgtc aagaaattgt tggacagata ctacttgtct      180

tttatcaacg acgttttgca ttccatcaag ctaaagaact tgaataacta catctctttg      240

ttcagaaaga agactagaac tgaaaaggaa aataaggaat tggaaaactt ggaaatcaac      300

ttgagaaagg aaattgctaa ggctttcaag ggtaatgaag gttacaagtc tttattcaag      360

aaagacatca ttgaaaccat tttgccagaa tttttggatg ataaggatga aattgctttg      420

gttaactctt tcaacggttt caccactgct ttcactggtt tcttcgacaa cagagaaaac      480

atgttctccg aggaagctaa atccacttct attgctttca gatgtatcaa cgaaaacttg      540

acccgttaca tctctaacat ggacattttt gaaaaggtcg acgccatctt tgacaagcac      600

gaagtccaag aaatcaagga aaagatctta aactccgact acgatgtcga agatttcttc      660

gaaggtgaat tcttcaactt tgttttaacc caagaaggta tcgatgtcta caacgccatt      720

atcggtggtt ttgtcactga atctggtgaa aagatcaagg gtttgaacga atacattaac      780

ttgtacaacc aaaagaccaa acaaaaattg ccaaagttca agccattgta caagcaagtt      840

ttgtctgaca gagaatcttt gtctttttac ggtgaagggt acacctctga cgaagaagtc      900

ttggaagtct tcagaaacac tttgaacaag aactctgaaa tcttctcctc catcaagaag      960

ttagaaaagt tgttcaagaa cttcgatgaa tactcttctg ctggtatctt cgttaagaac     1020

ggtccagcca tctctaccat ttctaaggat atctttggtg aatggaacgt cattagagac     1080

aaatggaacg ctgaatacga tgacatccat ttgaagaaaa aggctgttgt caccgaaaag     1140

tacgaagacg acagaagaaa atccttcaag aagatcggtt ccttctcctt ggaacaatta     1200

caagaatacg ccgatgccga tttgtccgtt gtcgaaaaat tgaaggaaat tattattcaa     1260

aaggttgatg aaatttacaa agtttacggt tcctctgaaa agttattcga tgctgatttc     1320

gtcttggaaa agtctttgaa gaagaacgac gctgttgtcg ctatcatgaa ggacttgttg     1380

gactctgtca aatctttcga aaactatatc aaggccttct tcggtgaagg taaggaaact     1440

aacagagatg aatccttcta cggtgacttt gtcttggctt acgatatttt gttgaaggtt     1500

gaccacatct acgatgccat cagaaactac gttactcaaa agccatactc taaggacaaa     1560

ttcaagttgt acttccaaaa cccacaattc atgggtggtt gggataagga caaggaaact     1620

gactacagag ctaccatttt gagatacggt tccaagtact acttggccat catggacaag     1680

aagtacgcca agtgtttgca aaagattgac aaggacgatg tcaacggtaa ctacgaaaag     1740

attaactaca agttgttgcc aggtccaaac aagatgttgc caaaggtttt cttctccaaa     1800

aagtggatgg cttactacaa cccatctgaa gacatccaaa agatctacaa gaacggtact     1860

ttcaaaaagg gtgacatgtt caacttaaac gactgtcaca agttgatcga cttcttcaag     1920

gactccatct ctagataccc aaaatggtcc aacgcttacg atttcaactt ctctgaaact     1980

gaaaaataca aggatattgc tggtttctac cgtgaagtcg aggaacaagg ttataaggtt     2040

tctttcgaat ccgcttctaa gaaagaagtt gacaaattag tcgaagaagg taagttgtac     2100

atgttccaaa tctacaacaa agatttctcc gacaagtctc acggtactcc aaacttgcac     2160

accatgtact tcaagttgct attcgatgaa aacaaccacg gtcaaatcag attgtctggt     2220

ggtgctgaat tgttcatgag acgtgcttct ctaaagaagg aagaattagt cgtccaccca     2280

gctaactctc caattgccaa caagaaccca gacaacccta agaagaccac cactttgtcc     2340

tacgacgttt acaaggacaa gagattctcc gaagaccaat acgaattgca cattccaatt     2400

gctatcaaca agtgtccaaa gaacatcttc aagatcaaca ctgaagtcag agttttgtta     2460

aagcacgatg acaaccctta cgttattggt atcgaccgtg gtgaaagaaa tttgttgtac     2520

attgttgttg ttgacggtaa gggtaacatc gttgaacaat actccttgaa cgaaatcatc     2580

aacaacttca acggtattag aatcaagact gattaccact ctttgttgga taagaaggaa     2640

aaggaacgtt ttgaagctcg tcaaaactgg acctctattg aaaacatcaa agaattgaag     2700

gctggttaca tcagtcaagt tgtccacaag atctgtgaat tggtcgagaa gtacgatgcc     2760

gttattgcct tggaagattt gaactctggt tttaagaact ctcgtgtcaa ggttgaaaag     2820

caagtctacc aaaagttcga aaagatgtta atcgacaaat tgaactacat ggttgacaag     2880

aaatccaacc catgtgctac cggtggtgct ttgaaaggtt accaaatcac caacaaattc     2940

gaatctttca aatctatgtc cactcaaaac gggttcatct tctacattcc agcttggttg     3000

acctccaaga tcgacccatc taccggtttc gttaacttgt tgaagaccaa gtacacttcc     3060

attgctgatt ccaagaagtt catctcttct ttcgacagaa tcatgtacgt tccagaagaa     3120

gacttgttcg aattcgcctt ggactataag aacttctcca gaaccgatgc tgactacatt     3180

aagaaatgga aattgtactc ctacggtaac agaatcagaa ttttcagaaa cccaaagaaa     3240

aacaacgttt tcgattggga agaagtttgt ttgacttctg cctacaagga attattcaac     3300

aaatacggta tcaactacca acaaggtgat atcagagctt tgttgtgtga acaatctgac     3360

aaggctttct actcttcctt catggctttg atgtccttga tgttgcaaat gagaaactcc     3420

atcactggta gaactgatgt cgacttcctc atttctccag ttaagaattc tgacggtatt     3480

ttctacgact ctagaaatta cgaagctcaa gaaaacgcta ttttgccaaa gaacgctgat     3540

gctaacggtg cttacaatat tgctagaaag gttttgtggg ctatcggtca attcaagaag     3600

gctgaagacg aaaagctaga caaggtcaag attgctattt ctaacaagga atggttggaa     3660

tacgctcaaa cctccgtcaa gcactccaga gctgatccaa agaagaagag aaaggtataa     3720


<210>  49
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify LbCpf1 
       expression cassette

<400>  49
cctcatagaa tattattctt cagtcactcg cttaaatact tatcaaaaat gtctaagttg       60


<210>  50
<211>  74
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify LbCpf1 
       expression cassette

<400>  50
cgtataatta tttgtgggaa cggctctaga aaagaaaact ttgcctttaa ctcctttata       60

cctttctctt cttc                                                         74


<210>  51
<211>  11322
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of vector pCSN067 encoding LbCpf1

<400>  51
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 tgtctaagtt ggaaaaattc accaactgtt actctttgtc     2940

taagactttg agattcaagg ccatcccagt tggtaagacc caagaaaaca tcgacaacaa     3000

gagactatta gttgaagatg aaaagagagc tgaagactac aagggtgtca agaaattgtt     3060

ggacagatac tacttgtctt ttatcaacga cgttttgcat tccatcaagc taaagaactt     3120

gaataactac atctctttgt tcagaaagaa gactagaact gaaaaggaaa ataaggaatt     3180

ggaaaacttg gaaatcaact tgagaaagga aattgctaag gctttcaagg gtaatgaagg     3240

ttacaagtct ttattcaaga aagacatcat tgaaaccatt ttgccagaat ttttggatga     3300

taaggatgaa attgctttgg ttaactcttt caacggtttc accactgctt tcactggttt     3360

cttcgacaac agagaaaaca tgttctccga ggaagctaaa tccacttcta ttgctttcag     3420

atgtatcaac gaaaacttga cccgttacat ctctaacatg gacatttttg aaaaggtcga     3480

cgccatcttt gacaagcacg aagtccaaga aatcaaggaa aagatcttaa actccgacta     3540

cgatgtcgaa gatttcttcg aaggtgaatt cttcaacttt gttttaaccc aagaaggtat     3600

cgatgtctac aacgccatta tcggtggttt tgtcactgaa tctggtgaaa agatcaaggg     3660

tttgaacgaa tacattaact tgtacaacca aaagaccaaa caaaaattgc caaagttcaa     3720

gccattgtac aagcaagttt tgtctgacag agaatctttg tctttttacg gtgaagggta     3780

cacctctgac gaagaagtct tggaagtctt cagaaacact ttgaacaaga actctgaaat     3840

cttctcctcc atcaagaagt tagaaaagtt gttcaagaac ttcgatgaat actcttctgc     3900

tggtatcttc gttaagaacg gtccagccat ctctaccatt tctaaggata tctttggtga     3960

atggaacgtc attagagaca aatggaacgc tgaatacgat gacatccatt tgaagaaaaa     4020

ggctgttgtc accgaaaagt acgaagacga cagaagaaaa tccttcaaga agatcggttc     4080

cttctccttg gaacaattac aagaatacgc cgatgccgat ttgtccgttg tcgaaaaatt     4140

gaaggaaatt attattcaaa aggttgatga aatttacaaa gtttacggtt cctctgaaaa     4200

gttattcgat gctgatttcg tcttggaaaa gtctttgaag aagaacgacg ctgttgtcgc     4260

tatcatgaag gacttgttgg actctgtcaa atctttcgaa aactatatca aggccttctt     4320

cggtgaaggt aaggaaacta acagagatga atccttctac ggtgactttg tcttggctta     4380

cgatattttg ttgaaggttg accacatcta cgatgccatc agaaactacg ttactcaaaa     4440

gccatactct aaggacaaat tcaagttgta cttccaaaac ccacaattca tgggtggttg     4500

ggataaggac aaggaaactg actacagagc taccattttg agatacggtt ccaagtacta     4560

cttggccatc atggacaaga agtacgccaa gtgtttgcaa aagattgaca aggacgatgt     4620

caacggtaac tacgaaaaga ttaactacaa gttgttgcca ggtccaaaca agatgttgcc     4680

aaaggttttc ttctccaaaa agtggatggc ttactacaac ccatctgaag acatccaaaa     4740

gatctacaag aacggtactt tcaaaaaggg tgacatgttc aacttaaacg actgtcacaa     4800

gttgatcgac ttcttcaagg actccatctc tagataccca aaatggtcca acgcttacga     4860

tttcaacttc tctgaaactg aaaaatacaa ggatattgct ggtttctacc gtgaagtcga     4920

ggaacaaggt tataaggttt ctttcgaatc cgcttctaag aaagaagttg acaaattagt     4980

cgaagaaggt aagttgtaca tgttccaaat ctacaacaaa gatttctccg acaagtctca     5040

cggtactcca aacttgcaca ccatgtactt caagttgcta ttcgatgaaa acaaccacgg     5100

tcaaatcaga ttgtctggtg gtgctgaatt gttcatgaga cgtgcttctc taaagaagga     5160

agaattagtc gtccacccag ctaactctcc aattgccaac aagaacccag acaaccctaa     5220

gaagaccacc actttgtcct acgacgttta caaggacaag agattctccg aagaccaata     5280

cgaattgcac attccaattg ctatcaacaa gtgtccaaag aacatcttca agatcaacac     5340

tgaagtcaga gttttgttaa agcacgatga caacccttac gttattggta tcgaccgtgg     5400

tgaaagaaat ttgttgtaca ttgttgttgt tgacggtaag ggtaacatcg ttgaacaata     5460

ctccttgaac gaaatcatca acaacttcaa cggtattaga atcaagactg attaccactc     5520

tttgttggat aagaaggaaa aggaacgttt tgaagctcgt caaaactgga cctctattga     5580

aaacatcaaa gaattgaagg ctggttacat cagtcaagtt gtccacaaga tctgtgaatt     5640

ggtcgagaag tacgatgccg ttattgcctt ggaagatttg aactctggtt ttaagaactc     5700

tcgtgtcaag gttgaaaagc aagtctacca aaagttcgaa aagatgttaa tcgacaaatt     5760

gaactacatg gttgacaaga aatccaaccc atgtgctacc ggtggtgctt tgaaaggtta     5820

ccaaatcacc aacaaattcg aatctttcaa atctatgtcc actcaaaacg ggttcatctt     5880

ctacattcca gcttggttga cctccaagat cgacccatct accggtttcg ttaacttgtt     5940

gaagaccaag tacacttcca ttgctgattc caagaagttc atctcttctt tcgacagaat     6000

catgtacgtt ccagaagaag acttgttcga attcgccttg gactataaga acttctccag     6060

aaccgatgct gactacatta agaaatggaa attgtactcc tacggtaaca gaatcagaat     6120

tttcagaaac ccaaagaaaa acaacgtttt cgattgggaa gaagtttgtt tgacttctgc     6180

ctacaaggaa ttattcaaca aatacggtat caactaccaa caaggtgata tcagagcttt     6240

gttgtgtgaa caatctgaca aggctttcta ctcttccttc atggctttga tgtccttgat     6300

gttgcaaatg agaaactcca tcactggtag aactgatgtc gacttcctca tttctccagt     6360

taagaattct gacggtattt tctacgactc tagaaattac gaagctcaag aaaacgctat     6420

tttgccaaag aacgctgatg ctaacggtgc ttacaatatt gctagaaagg ttttgtgggc     6480

tatcggtcaa ttcaagaagg ctgaagacga aaagctagac aaggtcaaga ttgctatttc     6540

taacaaggaa tggttggaat acgctcaaac ctccgtcaag cactccagag ctgatccaaa     6600

gaagaagaga aaggtataaa ggagttaaag gcaaagtttt cttttctaga gccgttccca     6660

caaataatta tacgtatatg cttcttttcg tttactatat atctatattt acaagccttt     6720

attcactgat gcaatttgtt tccaaatact tttttggaga tctcataact agatatcatg     6780

atggcgcaac ttggcgctat cttaattact ctggctgcca ggcccgtgta gagggccgca     6840

agaccttctg tacgccatat agtctctaag aacttgaaca agtttctaga cctattgccg     6900

cctttcggat cgctattgtt gcggccgcca gctgaagctt cgtacgctgc aggtcgacga     6960

attctaccgt tcgtataatg tatgctatac gaagttatag atctgtttag cttgcctcgt     7020

ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg     7080

acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc     7140

atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg     7200

gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc     7260

cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct     7320

ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata     7380

aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga     7440

tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat     7500

ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag     7560

cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc     7620

tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc     7680

gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat     7740

tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc     7800

ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt     7860

ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa     7920

agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc     7980

acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt     8040

cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc     8100

tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa     8160

attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt     8220

cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa     8280

tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag     8340

ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg     8400

ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctcata acttcgtata     8460

atgtatgcta tacgaacggt agaattcgaa tcagatccac tagtggccta tgcggccgcc     8520

accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta     8580

atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat     8640

aggagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt     8700

aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta     8760

atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc     8820

gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa     8880

ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa     8940

aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct     9000

ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac     9060

aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc     9120

gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc     9180

tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg     9240

tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga     9300

gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag     9360

cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta     9420

cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag     9480

agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg     9540

caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac     9600

ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc     9660

aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag     9720

tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc     9780

agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac     9840

gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc     9900

accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg     9960

tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag    10020

tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc    10080

acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac    10140

atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag    10200

aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac    10260

tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg    10320

agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc    10380

gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact    10440

ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg    10500

atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa    10560

tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt    10620

tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg    10680

tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg    10740

gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat    10800

aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag    10860

atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct    10920

ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc    10980

ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct    11040

tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta    11100

tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat    11160

acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga    11220

tacgaggcgc gtgtaagtta caggcaagcg atccgtccta agaaaccatt attatcatga    11280

cattaaccta taaaaatagg cgtatcacga ggccctttcg tc                       11322


<210>  52
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of direct repeat part of crRNA cassette of 
       LbCpf1

<400>  52
taatttctac taagtgtaga t                                                 21


<210>  53
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of guide sequence (genomic target sequence) 
       of INT1 for LbCpf1

<400>  53
ctggtgggag agaaagctta                                                   20


<210>  54
<211>  430
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the complete guide RNA cassette for 
       targeting LbCpf1 to the INT1 locus in the genome that contains 
       homology to vector backbone pRN1120 for homologous recombination

<400>  54
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat ctggtgggag agaaagctta      360

tttttttgtt ttttatgtct ggggggcccg gtacccagct tttgttccct ttagtgaggg      420

ttaattccga                                                             430


<210>  55
<211>  439
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-7 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       3' side

<400>  55
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct atttgtacta ctggtaaatt gccagttcca      360

tggccaacct tagtcactac tttaggttat ggtgcaatgt tttgctagat acccagatca      420

tatgaaacaa catgacttt                                                   439


<210>  56
<211>  489
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-8 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 3' side

<400>  56
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct ttgcccatcg aacgtacaag tactcctctg      360

ttctctcctt cctttgcttt atttgtacta ctggtaaatt gccagttcca tggccaacct      420

tagtcactac tttaggttat ggtgcaatgt tttgctagat acccagatca tatgaaacaa      480

catgacttt                                                              489


<210>  57
<211>  439
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-9 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       5' side

<400>  57
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt ctttgaaaag      120

ataatgtatg attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt      180

atatacaagg tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt      240

gggctagcgg taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt      300

ggtcaaacgc tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc      360

agtgaaagat aaatgatcta atttctacta agtgtagatc aatgttttgc tagataccct      420

ttttttgttt tttatgtct                                                   439


<210>  58
<211>  489
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-10 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 5' side

<400>  58
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt tgcccatcga      120

acgtacaagt actcctctgt tctctccttc ctttgctttt ctttgaaaag ataatgtatg      180

attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg      240

tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg      300

taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc      360

tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat      420

aaatgatcta atttctacta agtgtagatc aatgttttgc tagataccct ttttttgttt      480

tttatgtct                                                              489


<210>  59
<211>  459
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x 18 bp guide)

<400>  59
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagatactt tttttgtttt ttatgtcttt tgcaatgttt tgctagatac atttgtacta      360

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      420

tttgctagat acccagatca tatgaaacaa catgacttt                             459


<210>  60
<211>  463
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x 20 bp guide)

<400>  60
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct tttgcaatgt tttgctagat acccatttgt      360

actactggta aattgccagt tccatggcca accttagtca ctactttagg ttatggtgca      420

atgttttgct agatacccag atcatatgaa acaacatgac ttt                        463


<210>  61
<211>  459
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 18 bp guide)

<400>  61
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt      120

ttgctagata ctctttgaaa agataatgta tgattatgct ttcactcata tttatacaga      180

aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact      240

ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac      300

aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt      360

tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc taatttctac taagtgtaga      420

tcaatgtttt gctagatact ttttttgttt tttatgtct                             459


<210>  62
<211>  463
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 20 bp guide)

<400>  62
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt      120

ttgctagata ccctctttga aaagataatg tatgattatg ctttcactca tatttataca      180

gaaacttgat gttttctttc gagtatatac aaggtgatta catgtacgtt tgaagtacaa      240

ctctagattt tgtagtgccc tcttgggcta gcggtaaagg tgcgcatttt ttcacaccct      300

acaatgttct gttcaaaaga ttttggtcaa acgctgtaga agtgaaagtt ggtgcgcatg      360

tttcggcgtt cgaaacttct ccgcagtgaa agataaatga tctaatttct actaagtgta      420

gatcaatgtt ttgctagata cccttttttt gttttttatg tct                        463


<210>  63
<211>  430
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-7 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to INT1 and donor DNA on the 3' side

<400>  63
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat ctggtgggag      300

agaaagctta tttttttgtt ttttatgtct gtttcttttg gaaaacttca gtcgctcatt      360

attagaacca gggaggtcca ggcccggctg gtgggagaga aagcttatga agctggggtt      420

gcagatttgt                                                             430


<210>  64
<211>  480
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-8 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to INT1, connector A and donor DNA 
       on the 3'

<400>  64
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat ctggtgggag      300

agaaagctta tttttttgtt ttttatgtct ttgcccatcg aacgtacaag tactcctctg      360

ttctctcctt cctttgcttt gtttcttttg gaaaacttca gtcgctcatt attagaacca      420

gggaggtcca ggcccggctg gtgggagaga aagcttatga agctggggtt gcagatttgt      480


<210>  65

<400>  65
000

<210>  66

<400>  66
000

<210>  67
<211>  452
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to INT1, PAM and guide target 
       sequence and donor DNA on the 3' side (1 x 20 bp, 1x 18 bp guide)

<400>  67
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat ctggtgggag      300

agaaagctta tttttttgtt ttttatgtct tttgctggtg ggagagaaag ctgtttcttt      360

tggaaaactt cagtcgctca ttattagaac cagggaggtc caggcccggc tggtgggaga      420

gaaagcttat gaagctgggg ttgcagattt gt                                    452


<210>  68
<211>  454
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to INT1, PAM and guide target 
       sequence and donor DNA on the 3' side (2 x 20 bp guide)

<400>  68
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat ctggtgggag      300

agaaagctta tttttttgtt ttttatgtct tttgctggtg ggagagaaag cttagtttct      360

tttggaaaac ttcagtcgct cattattaga accagggagg tccaggcccg gctggtggga      420

gagaaagctt atgaagctgg ggttgcagat ttgt                                  454


<210>  69
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the guide sequence (genomic target) of the
       CTEC fragments targeting YFP by LbCpf1 in strain CSN010

<400>  69
caatgttttg ctagataccc                                                   20


<210>  70
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the guide sequence (genomic target) of the
       CTEC fragments targeting INT1 by LbCpf1 in strain CSN004

<400>  70
ctggtgggag agaaagctta                                                   20


<210>  71
<211>  109
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of YFP donor DNA that is part of CTEC 
       fragments for LbCpf1 mediated editing in strain CSN010

<400>  71
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgacttt                  109


<210>  72
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of INT donor DNA that is part of CTEC 
       fragments for LbCpf1 mediated editing in strain CSN004

<400>  72
gtttcttttg gaaaacttca gtcgctcatt attagaacca gggaggtcca ggcccggctg       60

gtgggagaga aagcttatga agctggggtt gcagatttgt                            100


<210>  73
<211>  330
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of complete guide RNA expression cassette for
       targeting LbCpf1 to the INT1 locus in the genome of CSN004

<400>  73
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat ctggtgggag      300

agaaagctta tttttttgtt ttttatgtct                                       330


<210>  74
<211>  330
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of complete guide RNA expression cassette for
       targeting LbCpf1 to the YFP expression cassette in the genome of 
       CSN010

<400>  74
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct                                       330


<210>  75
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 18 bp guide sequence (genomic target 
       sequence) for digestion of the CTEC fragment by LbCpf1 thereby 
       separating the INT1 donor DNA from the guide RNA expression 
       cassette

<400>  75
ctggtgggag agaaagct                                                     18


<210>  76
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 18 bp guide sequence (genomic target 
       sequence) for digestion of the CTEC fragment by LbCpf1 thereby 
       separating the YFP donor DNA from the guide RNA expression 
       cassette

<400>  76
caatgttttg ctagatac                                                     18


<210>  77
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20 bp guide sequence (genomic target 
       sequence) for digestion of the CTEC fragment by LbCpf1 thereby 
       separating the INT1 donor DNA from the guide RNA expression 
       cassette

<400>  77
ctggtgggag agaaagctta                                                   20


<210>  78
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20 bp guide sequence (genomic target 
       sequence) for digestion of the CTEC fragment by LbCpf1 thereby 
       separating the YFP donor DNA from the guide RNA expression 
       cassette

<400>  78
caatgttttg ctagataccc                                                   20


<210>  79
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 18 bp guide sequence (genomic target 
       sequence) including the PAM sequence for digestion of the CTEC 
       fragment by LbCpf1 thereby separating the INT1 donor DNA from the
       guide RNA expression cassette

<400>  79
tttgctggtg ggagagaaag ct                                                22


<210>  80
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20 bp guide sequence (genomic target 
       sequence) including the PAM sequence for digestion of the CTEC 
       fragment by LbCpf1 thereby separating the INT1 donor DNA from the
       guide RNA expression cassette

<400>  80
tttgctggtg ggagagaaag ctta                                              24


<210>  81
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 18 bp guide sequence (genomic target 
       sequence) including the PAM for digestion of the CTEC fragment by
       LbCpf1 thereby separating the YFP donor DNA from the guide RNA 
       expression cassette

<400>  81
tttgcaatgt tttgctagat ac                                                22


<210>  82
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20 bp guide sequence (genomic target 
       sequence) including the PAM sequence for digestion of the CTEC 
       fragment by LbCpf1 thereby separating the YFP donor DNA from the 
       guide RNA expression cassette

<400>  82
tttgcaatgt tttgctagat accc                                              24


<210>  83
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments having the YFP donor on the 5' side and a 20 bp guide 
       sequence for LbCpf1

<400>  83
agacataaaa aacaaaaaaa gggtatctag                                        30


<210>  84
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments having the YFP donor on the 5' side and a 18 bp guide 
       sequence for LbCpf1

<400>  84
agacataaaa aacaaaaaaa gtatctag                                          28


<210>  85
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify CTEC 
       fragments having the INT1 donor on the 5' side for LbCpf1 editing

<400>  85
gtttcttttg gaaaacttca gtcgc                                             25


<210>  86
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments having the INT1 donor on the 3' side for LbCpf1 editing

<400>  86
acaaatctgc aaccccagct tc                                                22


<210>  87
<211>  539
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-7 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       3' side, flanked by connector 5 sequence on the 5' side and 
       connector 3 on the 3' side

<400>  87
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagataccc      360

tttttttgtt ttttatgtct atttgtacta ctggtaaatt gccagttcca tggccaacct      420

tagtcactac tttaggttat ggtgcaatgt tttgctagat acccagatca tatgaaacaa      480

catgacttta gaaagcctgt atgcgaagcc acaatccttt ccaacagacc atactaagt       539


<210>  88
<211>  589
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-8 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 3' side, flanked by connector 5 sequence on the 
       5' side and connector 3 on the 3' side

<400>  88
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagataccc      360

tttttttgtt ttttatgtct ttgcccatcg aacgtacaag tactcctctg ttctctcctt      420

cctttgcttt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac      480

tttaggttat ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgacttta      540

gaaagcctgt atgcgaagcc acaatccttt ccaacagacc atactaagt                  589


<210>  89
<211>  539
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-9 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       5' side, flanked by connector 5 sequence on the 5' side and 
       connector 3 on the 3' side

<400>  89
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt ctttgaaaag ataatgtatg      180

attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg      240

tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg      300

taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc      360

tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat      420

aaatgatcta atttctacta agtgtagatc aatgttttgc tagataccct ttttttgttt      480

tttatgtcta gaaagcctgt atgcgaagcc acaatccttt ccaacagacc atactaagt       539


<210>  90
<211>  589
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-10 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 5' side, flanked by connector 5 sequence on the 
       5' side and connector 3 on the 3' side

<400>  90
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt tgcccatcga acgtacaagt      180

actcctctgt tctctccttc ctttgctttt ctttgaaaag ataatgtatg attatgcttt      240

cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg tgattacatg      300

tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg taaaggtgcg      360

cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg      420

aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat aaatgatcta      480

atttctacta agtgtagatc aatgttttgc tagataccct ttttttgttt tttatgtcta      540

gaaagcctgt atgcgaagcc acaatccttt ccaacagacc atactaagt                  589


<210>  91
<211>  559
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x 18 bp guide), 
       flanked by connector 5 sequence on the 5' side and connector 3 on
       

<400>  91
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagatactt      360

tttttgtttt ttatgtcttt tgcaatgttt tgctagatac atttgtacta ctggtaaatt      420

gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt tttgctagat      480

acccagatca tatgaaacaa catgacttta gaaagcctgt atgcgaagcc acaatccttt      540

ccaacagacc atactaagt                                                   559


<210>  92
<211>  563
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x  20 bp guide), 
       flanked by connector 5 sequence on the 5' side and connector 3 on
       

<400>  92
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagataccc      360

tttttttgtt ttttatgtct tttgcaatgt tttgctagat acccatttgt actactggta      420

aattgccagt tccatggcca accttagtca ctactttagg ttatggtgca atgttttgct      480

agatacccag atcatatgaa acaacatgac tttagaaagc ctgtatgcga agccacaatc      540

ctttccaaca gaccatacta agt                                              563


<210>  93
<211>  559
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 18 bp guide), 
       flanked by connector 5 on the 5' side and connector 3 on the 3' 
       

<400>  93
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt ttgctagata      180

ctctttgaaa agataatgta tgattatgct ttcactcata tttatacaga aacttgatgt      240

tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact ctagattttg      300

tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac aatgttctgt      360

tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt tcggcgttcg      420

aaacttctcc gcagtgaaag ataaatgatc taatttctac taagtgtaga tcaatgtttt      480

gctagatact ttttttgttt tttatgtcta gaaagcctgt atgcgaagcc acaatccttt      540

ccaacagacc atactaagt                                                   559


<210>  94
<211>  563
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 20 bp guide), 
       flanked by connector 5 on the 5' side and connector 3 on the 3' 
       

<400>  94
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt ttgctagata      180

ccctctttga aaagataatg tatgattatg ctttcactca tatttataca gaaacttgat      240

gttttctttc gagtatatac aaggtgatta catgtacgtt tgaagtacaa ctctagattt      300

tgtagtgccc tcttgggcta gcggtaaagg tgcgcatttt ttcacaccct acaatgttct      360

gttcaaaaga ttttggtcaa acgctgtaga agtgaaagtt ggtgcgcatg tttcggcgtt      420

cgaaacttct ccgcagtgaa agataaatga tctaatttct actaagtgta gatcaatgtt      480

ttgctagata cccttttttt gttttttatg tctagaaagc ctgtatgcga agccacaatc      540

ctttccaaca gaccatacta agt                                              563


<210>  95
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify CTEC 
       fragments with connector 5 on the 5' side

<400>  95
aagcgacttc caatcgcttt gcatatc                                           27


<210>  96
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify CTEC 
       fragments with connector 3 on the 3' side

<400>  96
cttagtatgg tctgttggaa aggattg                                           27


<210>  97
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of connector 5

<400>  97
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt                  50


<210>  98
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of connector 3

<400>  98
agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt                  50


<210>  99
<211>  489
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-7 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       3' side, flanked by connector 5 sequence on the 5' side

<400>  99
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagataccc      360

tttttttgtt ttttatgtct atttgtacta ctggtaaatt gccagttcca tggccaacct      420

tagtcactac tttaggttat ggtgcaatgt tttgctagat acccagatca tatgaaacaa      480

catgacttt                                                              489


<210>  100
<211>  539
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-8 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 3' side, flanked by connector 5 sequence on the 
       5' side

<400>  100
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagataccc      360

tttttttgtt ttttatgtct ttgcccatcg aacgtacaag tactcctctg ttctctcctt      420

cctttgcttt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac      480

tttaggttat ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgacttt       539


<210>  101
<211>  489
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-9 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       5' side, flanked by connector 5 sequence on the 5' side

<400>  101
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt ctttgaaaag ataatgtatg      180

attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg      240

tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg      300

taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc      360

tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat      420

aaatgatcta atttctacta agtgtagatc aatgttttgc tagataccct ttttttgttt      480

tttatgtct                                                              489


<210>  102
<211>  539
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-10 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 5' side, flanked by connector 5 sequence on the 
       5' side

<400>  102
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt tgcccatcga acgtacaagt      180

actcctctgt tctctccttc ctttgctttt ctttgaaaag ataatgtatg attatgcttt      240

cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg tgattacatg      300

tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg taaaggtgcg      360

cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg      420

aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat aaatgatcta      480

atttctacta agtgtagatc aatgttttgc tagataccct ttttttgttt tttatgtct       539


<210>  103
<211>  509
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x 18 bp guide), 
       flanked by connector 5 sequence on the 5' side

<400>  103
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagatactt      360

tttttgtttt ttatgtcttt tgcaatgttt tgctagatac atttgtacta ctggtaaatt      420

gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt tttgctagat      480

acccagatca tatgaaacaa catgacttt                                        509


<210>  104
<211>  513
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x  20 bp guide), 
       flanked by connector 5 sequence on the 5' side

<400>  104
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg ctagataccc      360

tttttttgtt ttttatgtct tttgcaatgt tttgctagat acccatttgt actactggta      420

aattgccagt tccatggcca accttagtca ctactttagg ttatggtgca atgttttgct      480

agatacccag atcatatgaa acaacatgac ttt                                   513


<210>  105
<211>  509
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 18 bp guide), 
       flanked by connector 5 sequence on the 5' side

<400>  105
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt ttgctagata      180

ctctttgaaa agataatgta tgattatgct ttcactcata tttatacaga aacttgatgt      240

tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact ctagattttg      300

tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac aatgttctgt      360

tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt tcggcgttcg      420

aaacttctcc gcagtgaaag ataaatgatc taatttctac taagtgtaga tcaatgtttt      480

gctagatact ttttttgttt tttatgtct                                        509


<210>  106
<211>  513
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 20 bp guide), 
       flanked by connector 5 sequence on the 5' side

<400>  106
aagcgacttc caatcgcttt gcatatccag taccacaccc acaggcgttt atttgtacta       60

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      120

tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt ttgctagata      180

ccctctttga aaagataatg tatgattatg ctttcactca tatttataca gaaacttgat      240

gttttctttc gagtatatac aaggtgatta catgtacgtt tgaagtacaa ctctagattt      300

tgtagtgccc tcttgggcta gcggtaaagg tgcgcatttt ttcacaccct acaatgttct      360

gttcaaaaga ttttggtcaa acgctgtaga agtgaaagtt ggtgcgcatg tttcggcgtt      420

cgaaacttct ccgcagtgaa agataaatga tctaatttct actaagtgta gatcaatgtt      480

ttgctagata cccttttttt gttttttatg tct                                   513


<210>  107
<211>  489
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-7 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       3' side, flanked by connector 3 sequence on the 3' side

<400>  107
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct atttgtacta ctggtaaatt gccagttcca      360

tggccaacct tagtcactac tttaggttat ggtgcaatgt tttgctagat acccagatca      420

tatgaaacaa catgacttta gaaagcctgt atgcgaagcc acaatccttt ccaacagacc      480

atactaagt                                                              489


<210>  108
<211>  539
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-8 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 3' side, flanked by connector 3 sequence on the 
       3' side

<400>  108
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct ttgcccatcg aacgtacaag tactcctctg      360

ttctctcctt cctttgcttt atttgtacta ctggtaaatt gccagttcca tggccaacct      420

tagtcactac tttaggttat ggtgcaatgt tttgctagat acccagatca tatgaaacaa      480

catgacttta gaaagcctgt atgcgaagcc acaatccttt ccaacagacc atactaagt       539


<210>  109
<211>  489
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-9 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene and donor DNA on the
       5' side, flanked by connector 3 sequence on the 3' side

<400>  109
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt ctttgaaaag      120

ataatgtatg attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt      180

atatacaagg tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt      240

gggctagcgg taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt      300

ggtcaaacgc tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc      360

agtgaaagat aaatgatcta atttctacta agtgtagatc aatgttttgc tagataccct      420

ttttttgttt tttatgtcta gaaagcctgt atgcgaagcc acaatccttt ccaacagacc      480

atactaagt                                                              489


<210>  110
<211>  539
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-10 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, connector A and 
       donor DNA on the 5' side, flanked by connector 3 sequence on the 
       3' side

<400>  110
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt tgcccatcga      120

acgtacaagt actcctctgt tctctccttc ctttgctttt ctttgaaaag ataatgtatg      180

attatgcttt cactcatatt tatacagaaa cttgatgttt tctttcgagt atatacaagg      240

tgattacatg tacgtttgaa gtacaactct agattttgta gtgccctctt gggctagcgg      300

taaaggtgcg cattttttca caccctacaa tgttctgttc aaaagatttt ggtcaaacgc      360

tgtagaagtg aaagttggtg cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat      420

aaatgatcta atttctacta agtgtagatc aatgttttgc tagataccct ttttttgttt      480

tttatgtcta gaaagcctgt atgcgaagcc acaatccttt ccaacagacc atactaagt       539


<210>  111
<211>  509
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x 18 bp guide), 
       flanked by connector 3 sequence on the 3' side

<400>  111
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagatactt tttttgtttt ttatgtcttt tgcaatgttt tgctagatac atttgtacta      360

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      420

tttgctagat acccagatca tatgaaacaa catgacttta gaaagcctgt atgcgaagcc      480

acaatccttt ccaacagacc atactaagt                                        509


<210>  112
<211>  509
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-11 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 3' side (2 x  20 bp guide), 
       flanked by connector 3 sequence on the 3' side

<400>  112
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagatactt tttttgtttt ttatgtcttt tgcaatgttt tgctagatac atttgtacta      360

ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtgcaatgt      420

tttgctagat acccagatca tatgaaacaa catgacttta gaaagcctgt atgcgaagcc      480

acaatccttt ccaacagacc atactaagt                                        509


<210>  113
<211>  513
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 18 bp guide), 
       flanked by connector 3 sequence on the 3' side

<400>  113
tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt       60

ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt      120

agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt      180

caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga      240

aacttctccg cagtgaaaga taaatgatct aatttctact aagtgtagat caatgttttg      300

ctagataccc tttttttgtt ttttatgtct tttgcaatgt tttgctagat acccatttgt      360

actactggta aattgccagt tccatggcca accttagtca ctactttagg ttatggtgca      420

atgttttgct agatacccag atcatatgaa acaacatgac tttagaaagc ctgtatgcga      480

agccacaatc ctttccaaca gaccatacta agt                                   513


<210>  114
<211>  513
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-12 comprising a guide RNA cassette 
       (crRNA) for targeting LbCpf1 to the YFP gene, PAM and guide 
       target sequence and donor DNA on the 5' side (2 x 20 bp guide), 
       flanked by connector 3 sequence on the 3' side

<400>  114
atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat       60

ggtgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt ttgcaatgtt      120

ttgctagata ccctctttga aaagataatg tatgattatg ctttcactca tatttataca      180

gaaacttgat gttttctttc gagtatatac aaggtgatta catgtacgtt tgaagtacaa      240

ctctagattt tgtagtgccc tcttgggcta gcggtaaagg tgcgcatttt ttcacaccct      300

acaatgttct gttcaaaaga ttttggtcaa acgctgtaga agtgaaagtt ggtgcgcatg      360

tttcggcgtt cgaaacttct ccgcagtgaa agataaatga tctaatttct actaagtgta      420

gatcaatgtt ttgctagata cccttttttt gttttttatg tctagaaagc ctgtatgcga      480

agccacaatc ctttccaaca gaccatacta agt                                   513


<210>  115
<211>  598
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for targeting Cas9 to the YFP gene and donor DNA of 60 
       bp, which encodes a frameshift, on the 3' side

<400>  115
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtcttt ccatggccaa ccttagtcac tactttagtt atggtttgca      480

atgttttgct agatacccga aaccttcgaa tccagccagc atgtcgacac ccacaagatg      540

tagtgcacgg ggggcccggt acccagcttt tgttcccttt agtgagggtt aattccga        598


<210>  116
<211>  618
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for targeting Cas9 to the YFP gene and donor DNA of 80 
       bp, which encodes a frameshift, on the 3' side

<400>  116
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtctaa attgccagtt ccatggccaa ccttagtcac tactttagtt      480

atggtttgca atgttttgct agatacccag atcatatgga aaccttcgaa tccagccagc      540

atgtcgacac ccacaagatg tagtgcacgg ggggcccggt acccagcttt tgttcccttt      600

agtgagggtt aattccga                                                    618


<210>  117
<211>  638
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for targeting Cas9 to the YFP gene and donor DNA of 100 
       bp, which encodes a frameshift, on the 3' side

<400>  117
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtctta ctactggtaa attgccagtt ccatggccaa ccttagtcac      480

tactttagtt atggtttgca atgttttgct agatacccag atcatatgaa acaacatgga      540

aaccttcgaa tccagccagc atgtcgacac ccacaagatg tagtgcacgg ggggcccggt      600

acccagcttt tgttcccttt agtgagggtt aattccga                              638


<210>  118
<211>  598
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for targeting Cas9 to the YFP gene and donor DNA of 60 
       bp, which encodes the full knock out of the YFP expression 
       cassette, on the 3' side

<400>  118
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtctcg tgctgagctc aacagtgccc aacccttgat tctttgtcat      480

cagacaactt gttgagtgga aaccttcgaa tccagccagc atgtcgacac ccacaagatg      540

tagtgcacgg ggggcccggt acccagcttt tgttcccttt agtgagggtt aattccga        598


<210>  119
<211>  618
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for targeting Cas9 to the YFP gene and donor DNA of 80 
       bp, which encodes the full knock out of the YFP expression 
       cassette, on the 3' side

<400>  119
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtctgc aatagttgcg tgctgagctc aacagtgccc aacccttgat      480

tctttgtcat cagacaactt gttgagtggt actaaaggga aaccttcgaa tccagccagc      540

atgtcgacac ccacaagatg tagtgcacgg ggggcccggt acccagcttt tgttcccttt      600

agtgagggtt aattccga                                                    618


<210>  120
<211>  638
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC-1 comprising a guide RNA cassette 
       (sgRNA) for targeting Cas9 to the YFP gene and donor DNA of 100 
       bp, which encodes the full knock out of the YFP expression 
       cassette, on the 3' side

<400>  120
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtctct tcatgccagc aatagttgcg tgctgagctc aacagtgccc      480

aacccttgat tctttgtcat cagacaactt gttgagtggt actaaaggag tgcttttcga      540

aaccttcgaa tccagccagc atgtcgacac ccacaagatg tagtgcacgg ggggcccggt      600

acccagcttt tgttcccttt agtgagggtt aattccga                              638


<210>  121
<211>  438
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the complete guide RNA expression cassette
       (sgRNA) for targeting Cas9 to the YFP expression cassette in the 
       genome of CSN009

<400>  121
cggagctagc atgcggccgc tctagaacta gtggatcccc cgggctgcag tctttgaaaa       60

gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag      120

tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct      180

tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt      240

tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg      300

cagtgaaaga taaatgatct tagtcactac tttaggttag ttttagagct agaaatagca      360

agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt      420

tttttgtttt ttatgtct                                                    438


<210>  122
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the guide sequence (genomic target) of the
       CTEC fragments targeting YFP by Cas9 in strain CSN009

<400>  122
ttagtcacta ctttaggtta                                                   20


<210>  123
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the donor DNA encoding a frameshift in the
       YFP gene, 60 bp

<400>  123
ttccatggcc aaccttagtc actactttag ttatggtttg caatgttttg ctagataccc       60


<210>  124
<211>  80
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the donor DNA encoding a frameshift in the
       YFP gene, 80 bp

<400>  124
aaattgccag ttccatggcc aaccttagtc actactttag ttatggtttg caatgttttg       60

ctagataccc agatcatatg                                                   80


<210>  125
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the donor DNA encoding a frameshift in the
       YFP gene, 100 bp

<400>  125
tactactggt aaattgccag ttccatggcc aaccttagtc actactttag ttatggtttg       60

caatgttttg ctagataccc agatcatatg aaacaacatg                            100


<210>  126
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the donor DNA encoding the knock out of 
       the YFP expression cassette, 60 bp

<400>  126
cgtgctgagc tcaacagtgc ccaacccttg attctttgtc atcagacaac ttgttgagtg       60


<210>  127
<211>  80
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the donor DNA encoding the knock out of 
       the YFP expression cassette, 80 bp

<400>  127
gcaatagttg cgtgctgagc tcaacagtgc ccaacccttg attctttgtc atcagacaac       60

ttgttgagtg gtactaaagg                                                   80


<210>  128
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the donor DNA encoding the knock out of 
       the YFP expression cassette, 100 bp

<400>  128
cttcatgcca gcaatagttg cgtgctgagc tcaacagtgc ccaacccttg attctttgtc       60

atcagacaac ttgttgagtg gtactaaagg agtgcttttc                            100


<210>  129
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer for amplification of 
       CTEC fragments (SEQ ID NO's: 115, 116, 117, 118, 119 and 120) 
       that are flanked by 50 bp sequences homologous to the linearized 
       pRN1120 vector backbone fragment (EcoRI and XhoI digested)

<400>  129
cggagctagc atgcggccg                                                    19


<210>  130
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer for amplification of 
       CTEC fragments (SEQ ID NO's: 115, 116, 117, 118, 119 and 120) 
       that are flanked by 50 bp sequences homologous to the linearized 
       pRN1120 vector backbone fragment (EcoRI and XhoI digested)

<400>  130
tcggaattaa ccctcactaa agg                                               23


<210>  131
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of connector F (CONF)

<400>  131
gaaaccttcg aatccagcca gcatgtcgac acccacaaga tgtagtgcac                  50


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

<220>
<223>  Nucleotide sequence of the wild-type genomic target (example 4)

<400>  132
ttagtcacta ctttaggtta                                                   20


<210>  133
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the modified genomic target (example 4)

<400>  133
ttagtcacta ctttagtta                                                    19


<210>  134
<211>  1587
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC DNA fragment 3, comprising a guide 
       RNA expression cassette (sgRNA) for targeting Cas9 to the GFP 
       gene and donor DNA of 100-bp, which encodes a 2 base modification
       in the PAM sequence, changing it from CGG to TAG, on the 3' side

<400>  134
acgaagaact gcggtcaggt gacacaactt tttccatctc agggtgtgtc gcgtgtgctt       60

catccaaact ttagttgggg ttcgggttcg cgcgagatga tcacgtgccc tgatttggtg      120

tcgtcccccg tcgcgctgcg cacgtgattt atttatttcc ggtggctgct gtctacgcgg      180

ggccttctct gcccttctgt ttcaaccttc gggcggttct cgtaaccagc agtagcaatc      240

catttcgaaa ctcaaagagc taaaaacgtt aaacctcagc agtcgctcga cgaatgggct      300

gcggttggga agcccacgag gcctatagcc agagcctcga gttgacagga gcccagacgc      360

cttttccaac ggcaactttt atataaaatg gcaatgtatt catgcaattg cggccgtgtc      420

aggttggaga cactggacca cactctccat tgcttcctga ggagatggat cattgctagt      480

gcatctacgc gcagcaatcc cgcaagctcg acaaccgtag atgggctttg gtgggccaat      540

caattacgca acccgcacgt taaattgtat gaggaaggaa ggccacggta caaagtgggt      600

ggtcttcacc cagtggttgt tggtggcgtc atgcagacca tgcattgggg atagcacagg      660

gttggggtgt cttgtggact caatgggtga aaggagatgg aaaagggcgg tgaaaagtgg      720

tagaatcgaa atccctgacg tcaatttata aagtaaaatg cgtttctgcc attttgctcc      780

cctccttctt tcgcaatcgc ctccccaaaa gttgtcgtgg cagtacacat gcttgcatac      840

aatgaagcta atccggcttg ctcagtagtt gctatatcca ggcatggtgt gaaacccctc      900

aaagtatata taggagcggt gagccccagt ctggggtctt ttctctccat ctcaaaacta      960

ctttctcaca atggtattgc tgatgagtcc gtgaggacga aacgagtaag ctcgtccaat     1020

acccttaagc tcgattgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1080

atcaacttga aaaagtggca ccgagtcggt gcttttggcc ggcatggtcc cagcctcctc     1140

gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggactaaact tcgagctaat     1200

ccagtagctt acgttaccca ggggcaggtc aactggctag ccacgagtct gtcccaggtc     1260

gcaatttagt gtaataaaca atatatatat tgagtctaaa gggaattgta gctattgtga     1320

ttgtgtgatt ttcgtcttgc tggttcttat tgtgtcccat tcgtttcatc ctgatgagga     1380

cccctggaac cggtgttttc ttagtctctg caatcgctag tcttgttgct atgacagttg     1440

cgtcgacact attcaggtca tctatcggtt attctgatat tataatatcc agcttgtgac     1500

cgagaatgtt accatcctcc ttgaaatcaa tacccttaag ctcgatttag ttaacgaggg     1560

tatcaccctc aaacttaacc tcagctc                                         1587


<210>  135
<211>  1587
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC DNA fragment 4, comprising a guide 
       RNA expression cassette (sgRNA) for targeting Cas9 to the GFP 
       gene and donor DNA of 100-bp, which encodes a silent mutation in 
       the GFP gene by changing the PAM sequence from CGG to CGA

<400>  135
acgaagaact gcggtcaggt gacacaactt tttccatctc agggtgtgtc gcgtgtgctt       60

catccaaact ttagttgggg ttcgggttcg cgcgagatga tcacgtgccc tgatttggtg      120

tcgtcccccg tcgcgctgcg cacgtgattt atttatttcc ggtggctgct gtctacgcgg      180

ggccttctct gcccttctgt ttcaaccttc gggcggttct cgtaaccagc agtagcaatc      240

catttcgaaa ctcaaagagc taaaaacgtt aaacctcagc agtcgctcga cgaatgggct      300

gcggttggga agcccacgag gcctatagcc agagcctcga gttgacagga gcccagacgc      360

cttttccaac ggcaactttt atataaaatg gcaatgtatt catgcaattg cggccgtgtc      420

aggttggaga cactggacca cactctccat tgcttcctga ggagatggat cattgctagt      480

gcatctacgc gcagcaatcc cgcaagctcg acaaccgtag atgggctttg gtgggccaat      540

caattacgca acccgcacgt taaattgtat gaggaaggaa ggccacggta caaagtgggt      600

ggtcttcacc cagtggttgt tggtggcgtc atgcagacca tgcattgggg atagcacagg      660

gttggggtgt cttgtggact caatgggtga aaggagatgg aaaagggcgg tgaaaagtgg      720

tagaatcgaa atccctgacg tcaatttata aagtaaaatg cgtttctgcc attttgctcc      780

cctccttctt tcgcaatcgc ctccccaaaa gttgtcgtgg cagtacacat gcttgcatac      840

aatgaagcta atccggcttg ctcagtagtt gctatatcca ggcatggtgt gaaacccctc      900

aaagtatata taggagcggt gagccccagt ctggggtctt ttctctccat ctcaaaacta      960

ctttctcaca atggtattgc tgatgagtcc gtgaggacga aacgagtaag ctcgtccaat     1020

acccttaagc tcgattgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1080

atcaacttga aaaagtggca ccgagtcggt gcttttggcc ggcatggtcc cagcctcctc     1140

gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggactaaact tcgagctaat     1200

ccagtagctt acgttaccca ggggcaggtc aactggctag ccacgagtct gtcccaggtc     1260

gcaatttagt gtaataaaca atatatatat tgagtctaaa gggaattgta gctattgtga     1320

ttgtgtgatt ttcgtcttgc tggttcttat tgtgtcccat tcgtttcatc ctgatgagga     1380

cccctggaac cggtgttttc ttagtctctg caatcgctag tcttgttgct atgacagttg     1440

cgtcgacact attcaggtca tctatcggtt attctgatat tataatactc cagcttgtga     1500

ccgagaatgt taccatcctc ctagaaatca atacccttaa gctcgattcg attaacgagg     1560

gtatcaccct caaacttaac ctcagct                                         1587


<210>  136
<211>  973
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Yarrowia Yl_HYPO promoter

<400>  136
acgaagaact gcggtcaggt gacacaactt tttccatctc agggtgtgtc gcgtgtgctt       60

catccaaact ttagttgggg ttcgggttcg cgcgagatga tcacgtgccc tgatttggtg      120

tcgtcccccg tcgcgctgcg cacgtgattt atttatttcc ggtggctgct gtctacgcgg      180

ggccttctct gcccttctgt ttcaaccttc gggcggttct cgtaaccagc agtagcaatc      240

catttcgaaa ctcaaagagc taaaaacgtt aaacctcagc agtcgctcga cgaatgggct      300

gcggttggga agcccacgag gcctatagcc agagcctcga gttgacagga gcccagacgc      360

cttttccaac ggcaactttt atataaaatg gcaatgtatt catgcaattg cggccgtgtc      420

aggttggaga cactggacca cactctccat tgcttcctga ggagatggat cattgctagt      480

gcatctacgc gcagcaatcc cgcaagctcg acaaccgtag atgggctttg gtgggccaat      540

caattacgca acccgcacgt taaattgtat gaggaaggaa ggccacggta caaagtgggt      600

ggtcttcacc cagtggttgt tggtggcgtc atgcagacca tgcattgggg atagcacagg      660

gttggggtgt cttgtggact caatgggtga aaggagatgg aaaagggcgg tgaaaagtgg      720

tagaatcgaa atccctgacg tcaatttata aagtaaaatg cgtttctgcc attttgctcc      780

cctccttctt tcgcaatcgc ctccccaaaa gttgtcgtgg cagtacacat gcttgcatac      840

aatgaagcta atccggcttg ctcagtagtt gctatatcca ggcatggtgt gaaacccctc      900

aaagtatata taggagcggt gagccccagt ctggggtctt ttctctccat ctcaaaacta      960

ctttctcaca atg                                                         973


<210>  137
<211>  6
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 6-bp inverted repeat of the guide 
       sequence of the GFP gene

<400>  137
gtattg                                                                   6


<210>  138
<211>  37
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the HH ribozyme

<400>  138
ctgatgagtc cgtgaggacg aaacgagtaa gctcgtc                                37


<210>  139
<211>  148
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the HDV ribozyme

<400>  139
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtgctttt ggccggcatg gtcccagcct cctcgctggc gccggctggg      120

caacatgctt cggcatggcg aatgggac                                         148


<210>  140
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20-bp genomic target sequence of the 
       GFP gene

<400>  140
caataccctt aagctcgatt                                                   20


<210>  141
<211>  303
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the Yarrowia Yl_PGM terminator

<400>  141
taaacttcga gctaatccag tagcttacgt tacccagggg caggtcaact ggctagccac       60

gagtctgtcc caggtcgcaa tttagtgtaa taaacaatat atatattgag tctaaaggga      120

attgtagcta ttgtgattgt gtgattttcg tcttgctggt tcttattgtg tcccattcgt      180

ttcatcctga tgaggacccc tggaaccggt gttttcttag tctctgcaat cgctagtctt      240

gttgctatga cagttgcgtc gacactattc aggtcatcta tcggttattc tgatattata      300

ata                                                                    303


<210>  142
<211>  1487
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of guide-RNA expression cassette (sgRNA) 
       targeting the GFP gene

<400>  142
acgaagaact gcggtcaggt gacacaactt tttccatctc agggtgtgtc gcgtgtgctt       60

catccaaact ttagttgggg ttcgggttcg cgcgagatga tcacgtgccc tgatttggtg      120

tcgtcccccg tcgcgctgcg cacgtgattt atttatttcc ggtggctgct gtctacgcgg      180

ggccttctct gcccttctgt ttcaaccttc gggcggttct cgtaaccagc agtagcaatc      240

catttcgaaa ctcaaagagc taaaaacgtt aaacctcagc agtcgctcga cgaatgggct      300

gcggttggga agcccacgag gcctatagcc agagcctcga gttgacagga gcccagacgc      360

cttttccaac ggcaactttt atataaaatg gcaatgtatt catgcaattg cggccgtgtc      420

aggttggaga cactggacca cactctccat tgcttcctga ggagatggat cattgctagt      480

gcatctacgc gcagcaatcc cgcaagctcg acaaccgtag atgggctttg gtgggccaat      540

caattacgca acccgcacgt taaattgtat gaggaaggaa ggccacggta caaagtgggt      600

ggtcttcacc cagtggttgt tggtggcgtc atgcagacca tgcattgggg atagcacagg      660

gttggggtgt cttgtggact caatgggtga aaggagatgg aaaagggcgg tgaaaagtgg      720

tagaatcgaa atccctgacg tcaatttata aagtaaaatg cgtttctgcc attttgctcc      780

cctccttctt tcgcaatcgc ctccccaaaa gttgtcgtgg cagtacacat gcttgcatac      840

aatgaagcta atccggcttg ctcagtagtt gctatatcca ggcatggtgt gaaacccctc      900

aaagtatata taggagcggt gagccccagt ctggggtctt ttctctccat ctcaaaacta      960

ctttctcaca atggtattgc tgatgagtcc gtgaggacga aacgagtaag ctcgtccaat     1020

acccttaagc tcgattgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1080

atcaacttga aaaagtggca ccgagtcggt gcttttggcc ggcatggtcc cagcctcctc     1140

gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggactaaact tcgagctaat     1200

ccagtagctt acgttaccca ggggcaggtc aactggctag ccacgagtct gtcccaggtc     1260

gcaatttagt gtaataaaca atatatatat tgagtctaaa gggaattgta gctattgtga     1320

ttgtgtgatt ttcgtcttgc tggttcttat tgtgtcccat tcgtttcatc ctgatgagga     1380

cccctggaac cggtgttttc ttagtctctg caatcgctag tcttgttgct atgacagttg     1440

cgtcgacact attcaggtca tctatcggtt attctgatat tataata                   1487


<210>  143
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of 100-bp donor DNA of CTEC DNA fragment 1

<400>  143
gggaaacatg tcctggactt acaacttgct tcgctcttga tcttcggata gtagtataag       60

tgtgtgtgtt ggtgctaata atccgtcctc tccacccctt                            100


<210>  144
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of 100-bp donor DNA of CTEC DNA fragment 2

<400>  144
tccagcttgt gaccgagaat gttaccatcc tccttgaaat caataccctt aagctcgatt       60

cgttaacgag ggtatcaccc tcaaacttaa cctcagctcg                            100


<210>  145
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of 100-bp donor DNA of CTEC DNA fragment 3

<400>  145
tccagcttgt gaccgagaat gttaccatcc tccttgaaat caataccctt aagctcgatt       60

tagttaacga gggtatcacc ctcaaactta acctcagctc                            100


<210>  146
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of 100-bp donor DNA of CTEC DNA fragment 4

<400>  146
ctccagcttg tgaccgagaa tgttaccatc ctcctagaaa tcaataccct taagctcgat       60

tcgattaacg agggtatcac cctcaaactt aacctcagct                            100


<210>  147
<211>  11606
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of plasmid MB7452

<400>  147
cgcgtggatc gccggtgcgt tgacgttggt gacctccagc cagaggtgcc cggcgccccg       60

ctcgccggcg aactccgtcg cgagccccat caacgcgcgc ccgaccccgt gcccccggtg      120

ctccggggcg acctcgatgt cctcgacggt cagccggcgg ttccacgccg agtacgagat      180

gaccacgaag cccgccaggt cgccgtcgtc cccgtacgcg acgaacgtcc gggagtccgg      240

gtcgccgtcc tccccgtcgt ccgattcgtc gtccgattcg tcgtcgggga acaccttggt      300

caggggcggg tccaccggca cctcccgcag ggtgaagccg tccccggtgg cggtgacgcg      360

gaagacggtg tcggtggtga aggacccatc cagtgcctcg atggcctcgg cgtcccccgg      420

gacactggtg cggtaccggt aagccgtgtc gtcaagagtg gtcatttttg tgtctaggtg      480

tttgtgtttg gactgcgatc agtgaagaaa agaagaggaa aaattgtgca agaaattttg      540

ctttcaagac ttggctgatg cagcagggta actctgggac acagacctat gtttgtggtt      600

aaactcaatg cacgtggtac gtgcgtggag cgcttaccca tccaagggtg tggacatgga      660

accgacggtc cgtggagttg tgtaatgtca ttttggcgac tcttgaagca aggctataaa      720

aaaattgtgt ggcttgagtc ttatcgagct cggtcactac aagagttaat cttcctgtct      780

caggcagaca ggtcaggcag ggttactttt gggtgtgctg taactcactg tatggccgtt      840

agtgcgcata gacgttgtac atactggacc gaattgtagc gtgctcaata gggccaataa      900

agctattgta gggatccgaa ttttcagaac ctaatttatc tgttacccgg cctgtggctc      960

gcacagctta aaaatggtca aactttcccc ttcttgtctt tttttcctca cattcatcag     1020

gttcttgtct tgatctttca agtgagtatt aattaccgac cttggttctt cattgggaga     1080

gcattggaag ccgtggtgca gcaaccacaa aacggttctt ccccttcgat accttcttgc     1140

ctgcctttca atacaagtcg gctcgattag cggtggtcgc ccccgccagc ggagaacatg     1200

gaactaaccc agaatgagag ctaagtggag aaagaagaga gtcagacgac tcaagcgaaa     1260

gcgccgcaag gtccgagctc gatccaaata agcggttttt aacggagatt taacactaaa     1320

tcgaagaact tttcccgttt catttgcgaa tgagctcgtt aacaaaatcc cccagttttt     1380

ttatccagct gtaaggattg acattagtaa tgaattattg tttggtatat ttaaatctgt     1440

agttcctttc tgtccgtgtc ggcaactgtc gtactcgtga tttacttgta ttgacgaata     1500

cttactgtag cgcactctgc tgctactggt cgtaaggatg tgctatttcg gtgtatggtg     1560

ggttttttgg gggtcggaac cgaagactgt tacacgggca cggctcgttg tgtacacgca     1620

cagagctctt gcgagtcatg ttgtagctag ctcgtcgtgt tcaggaactg ttcgatggtt     1680

cggagagagt cgccgcccag aacatacgcg caccgatgtc agcagacagc cttattacaa     1740

gtatattcaa gcaagtatat ccgtagggtg cgggtgattt ggatctaagg ttcgtactca     1800

acactcacga gcagcttgcc tatgttacat ccttttatca gacataacat aattggagtt     1860

tacttacaca cggggtgtac ctgtatgagc accacctaca attgtagcac tggtacttgt     1920

acaaagaatt tattcgtacg aatcacaggg acggccgccc tcaccgaacc agcgaatacc     1980

tcagcggtcc cctgcagtga ctcaacaaag cgatatgaac atcttgcgat ggtatcctgc     2040

tgatagtttt tactgtacaa acacctgtgt agctccttct agcattttta agttattcac     2100

acctcaaggg gagggataaa ttaaataaat tccaaaagcg aagatcgaga aactaaatta     2160

aaattccaaa aacgaagttg gaacacaacc ccccgaaaaa aaacaacaaa caaaaaaccc     2220

aacaaaataa acaaaaacaa aataaatata taactaccag tatctgacta aaagttcaaa     2280

tactcgtact tacaacaaat agaaatgagc cggccaaaat tctgcagaaa aaaatttcaa     2340

acaagtactg gtataattaa attaaaaaac acatcaaagt atcataacgt tagttatttt     2400

attttattta ataaaagaaa acaacaagat gggctcaaaa ctttcaactt atacgataca     2460

taccaaataa caatttagta tttatctaag tgcttttcgt agataatgga atacaaatgg     2520

atatccagag tatacacatg gatagtatac actgacacga caattctgta tctctttatg     2580

ttaactactg tgaggcatta aatagagctt gatatataaa atgttacatt tcacagtctg     2640

aacttttgca gattacctaa tttggtaaga tattaattat gaactgaaag ttgatggcat     2700

ccctaaattt gatgaaagat gaaattgtaa atgaggtggt aaaagagcta cagtcgtttt     2760

gttttgagat accatcatct ctaacgaaat atctattaaa aatctcagtg tgatcatgag     2820

tcattgccat cctggaaaat gtcatcatgg ctgatatttc taactgttta cttgagataa     2880

atatatattt acaagaactt cccttgaaat taatttagat ataaaatgtt tgcgggcaag     2940

ttactacgag gaataaatta tatctagagg ttccgcttcc tcgctcactg actcgctgcg     3000

ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc     3060

cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag     3120

gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca     3180

tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca     3240

ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg     3300

atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag     3360

gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt     3420

tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca     3480

cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg     3540

cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt     3600

tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc     3660

cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg     3720

cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg     3780

gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta     3840

gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg     3900

gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg     3960

ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc     4020

atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc     4080

agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc     4140

ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag     4200

tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat     4260

ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg     4320

caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt     4380

gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag     4440

atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg     4500

accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt     4560

aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct     4620

gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac     4680

tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat     4740

aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat     4800

ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca     4860

aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat     4920

tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tggcctagga     4980

agcgacttcc aatcgctttg catatccagt accacaccca caggcgtttg tgctactcta     5040

ctgatagcaa tagatgcgtc ataattggtt ggcccgctga gcctccacag gatactattg     5100

cacataccct ggtcatgtgc agatcagctc atttgtggag actctggagt aacttagacg     5160

acgcctggtt caattgccgc aatgtgcgcc cacgcagata atgtattgag gggtggagcg     5220

cctcttgggg acttgctgta cttgtacggg atattaaacg cactcagcaa gaccatgacg     5280

taaaacacac ctactgtacg atacgtactg taggtattgt actcgtaccc ggtactacaa     5340

atagtacgat actatacgga gtgtatttgt accttgatat acgactggcg gagtgaagag     5400

aaggagttga acaagaccag atggggatat cagccccagt gctttgtatt acaagtacga     5460

gtacttaata gatactgtaa ggctattgat acggatggca gtaagtcatt gagtaagcaa     5520

ttgtggccca gcatctcccc tacgtacttg taccataccc catggagaca ccaatggtct     5580

ttcacgcaca ctgtcgtgtg ctgtatcgca gaatcgggtg tccaaccaaa tgccgttacc     5640

cccacgtcac agccgataga cagatacacc atcaatacca gcaggttgta tcatgcggtt     5700

ggctgaaggt aagctgattg gtctaaaaac tgtagctgtc ctaattcaac gagcgctatt     5760

tggggccaac cacctcggcc aagcggcctt taatctgcgt gccccagagg cgtctaatga     5820

ggctctggcc gccactgtag gagtgtttct ctgtgcgcac acgcagtttt gagtttgggc     5880

gactttccct ttttcccaat tgcgtacaca cacagctccg agctaagcgc tgtccttgaa     5940

ccttctccct cttttccctc tttttctctt ccccttcccc tcctccacat taaggccaaa     6000

tcctgaattg caccaactag tacaacgaca acaatggaca agaagtactc catcggtttg     6060

gacattggta ctaactctgt cggctgggcc gtcatcaccg acgagtacaa ggttccctcc     6120

aagaagttca aggtccttgg caacaccgac cgacactcta tcaagaagaa cctgatcggt     6180

gctctgctgt tcgactctgg cgagactgcc gaggccaccc gactgaagcg aaccgctcga     6240

cgccgataca cccgacgaaa gaaccgaatc tgttacctcc aggagatctt cagcaacgag     6300

atggctaagg tcgacgactc cttcttccac cgactcgagg agtctttcct ggtcgaagag     6360

gataagaagc acgagcgaca ccccatcttc ggcaacattg ttgatgaggt tgcctaccat     6420

gagaagtacc ccaccatcta ccacctccga aagaagctcg tcgactccac tgacaaggct     6480

gacctccgac tcatctacct tgctctcgcc cacatgatca agttccgagg tcacttcctc     6540

attgagggtg atctcaaccc cgacaactcc gacgttgaca agctgttcat ccagctcgtc     6600

cagacctaca accagctctt tgaggagaac cctatcaacg cttctggtgt tgacgccaag     6660

gccattctct ccgcccgact ctctaagtcc cgacgactcg agaacctcat tgcccagctg     6720

cccggcgaga agaagaacgg cctcttcggt aacctgattg ctctctctct tggtctgacc     6780

cccaacttca agtccaactt tgacctcgcc gaggacgcca agctccagct gtccaaggac     6840

acctacgatg acgatctgga caacctcctg gcccagatcg gtgaccagta cgccgatctc     6900

ttccttgccg ccaagaacct ctccgacgcc atcctgctct ccgacatcct ccgagtcaac     6960

accgagatta ccaaggctcc tctgtctgcc tctatgatca agcgatacga cgagcaccac     7020

caggatctca ctcttctcaa ggctctcgtc cgacagcagc tccccgagaa gtacaaggag     7080

attttctttg accagtccaa gaacggttac gctggctaca ttgacggtgg tgcttcccag     7140

gaagagtttt acaagttcat caagcctatt ctggagaaga tggacggtac cgaggagctg     7200

ctcgtcaagc tcaaccgaga ggacctcctt cgaaagcagc gaaccttcga taacggctcc     7260

atcccccacc agatccacct gggtgagctc cacgccattc tccgaagaca agaggacttc     7320

taccccttcc taaaggataa ccgagagaag atcgagaaga ttctcacctt ccgaatcccc     7380

tactacgtcg gtcccctcgc tcgaggtaac tcccgatttg cttggatgac ccgaaagtcc     7440

gaggagacta tcaccccctg gaactttgaa gaggtagtcg acaagggtgc ctccgcccag     7500

tctttcattg agcggatgac caacttcgat aagaacctcc ccaacgagaa ggtccttccc     7560

aagcactctc tcctctacga gtacttcacc gtctacaacg agctgaccaa ggtcaagtac     7620

gttaccgagg gcatgcgaaa gcccgctttc ctctctggtg agcagaagaa ggccattgtc     7680

gacctcctgt tcaagactaa ccgaaaagtc accgtcaagc agctcaagga agactacttc     7740

aagaagattg agtgcttcga ctccgtcgag atttccggtg tcgaggaccg attcaacgcc     7800

tccctcggca cctaccacga tcttctgaag atcatcaagg acaaggactt tcttgataac     7860

gaggagaacg aggacattct cgaggacatc gtcctcaccc tcaccctttt cgaggatcga     7920

gagatgatcg aggagcgact caagacctac gcccatctct tcgacgacaa ggtcatgaag     7980

caactcaagc gacgacgata cactggctgg ggccgacttt cccgaaagct catcaacggc     8040

atccgagaca agcagtctgg caagaccatc ctggacttcc tgaagtccga cggtttcgcc     8100

aaccgaaact tcatgcagct catccacgac gactctctta ccttcaaaga ggatatccag     8160

aaggcccagg tttctggcca gggcgactcc ctccacgagc acattgccaa cctcgccgga     8220

tcccccgcca tcaaaaaggg tatcctccag accgtcaagg ttgtcgacga actcgtgaag     8280

gtcatgggcc gacacaagcc cgagaacatc gttatcgaga tggcccgaga gaaccagacc     8340

acccagaagg gtcagaagaa ctcccgagag cgaatgaagc gaatcgaaga gggtatcaag     8400

gagctcggtt cccagattct caaggagcac cccgtcgaga acacccagct ccagaacgag     8460

aaactctacc tgtactacct ccagaatggc cgagacatgt acgttgacca ggagctcgac     8520

atcaaccgac tctccgacta cgacgtcgac cacattgttc ctcagtcctt cctcaaggac     8580

gactccatcg acaacaaggt tctgacccga tctgacaaga accgaggtaa gtccgacaac     8640

gttccctccg aagaggtcgt taagaagatg aagaactact ggcgacagct tctcaacgcc     8700

aaactgatca cccagcgaaa gtttgacaac ctcaccaagg ccgagcgagg tggtctgtcc     8760

gagctggaca aggccggctt cattaagcga cagctggtcg agactcgaca gatcaccaag     8820

cacgtcgccc agatcctcga ctcccgaatg aacaccaagt acgacgagaa cgacaagctc     8880

atccgggagg tcaaggtcat caccctgaag tctaagcttg tctccgactt ccgaaaggac     8940

ttccagttct acaaggtccg agagatcaac aactaccacc acgcccacga cgcctacctc     9000

aacgccgttg ttggtaccgc cctcatcaag aagtatccca agctcgagtc cgagttcgtt     9060

tacggcgact acaaggttta cgatgtccga aagatgattg ccaagtccga gcaggagatc     9120

ggtaaggcca ccgccaagta ctttttctac tccaacatca tgaatttctt caagaccgag     9180

atcactctcg ccaacggtga gattcgaaag cgacccctga ttgagactaa tggtgagact     9240

ggtgagatcg tctgggataa gggccgagac ttcgccaccg tccgaaaggt cctgtccatg     9300

ccccaggtca acattgtcaa gaagaccgag gtccagaccg gtggcttctc caaggagtcc     9360

attctcccca agcgaaactc cgacaaactc atcgcccgta agaaggactg ggatccgaag     9420

aagtacggtg gtttcgattc tcccaccgtt gcctactccg tcctcgttgt tgctaaagtc     9480

gagaagggta agtctaagaa actcaagtcc gtgaaggagc tactcggtat caccatcatg     9540

gagcgatctt cttttgagaa gaaccccatt gacttcctcg aggccaaggg ttacaaagag     9600

gtcaagaagg acctgattat caagctgccc aagtactccc tctttgagct cgagaacggc     9660

cgaaagcgaa tgctggcttc cgctggtgag ctgcagaagg gcaacgagct cgctctgccc     9720

tccaagtacg tcaacttcct ctacctggcc tcccactacg agaagctcaa gggctccccc     9780

gaggacaacg agcagaagca gctgttcgtt gagcagcaca agcactacct cgacgagatc     9840

atcgagcaga tctccgagtt ctccaagcga gtcatcctcg ctgacgccaa ccttgataag     9900

gttctctctg cttacaacaa gcaccgggac aagcccatcc gagagcaggc cgagaatatc     9960

atccacctct tcactctcac caacctcggc gctcctgctg ccttcaagta cttcgacacc    10020

accattgacc gaaagaggta cacctccacc aaggaagtcc tcgacgccac cctgatccac    10080

cagtccatca ccggcctcta cgaaacccga atcgacctct cccagctcgg cggtgactct    10140

cgagccgacc ccaagaagaa gcgaaaagtc taaatatccg aagatcaaga gcgaagcaag    10200

ttgtaagtcc aggacatgtt tcccgcccac gcgagtgatt tataacacct ctcttttttg    10260

acacccgctc gccttgaaat tcatgtcaca taaattatag tcaacgacgt ttgaataact    10320

tgtcttgtag ttcgatgatg atcatatgat tacattaata gtaattactg tatttgatat    10380

atatactaat tacaatagta catattagaa catacaatag ttagtgccgt gaagtggctt    10440

aaaataccgc gagtcgatta cgtaatatta ttacctcttg cccatcgaac gtacaagtac    10500

tcctctgttc tctccttcct ttgctttgtg cacgaagaac tgcggtcagg tgacacaact    10560

ttttccatct cagggtgtgt cgcgtgtgct tcatccaaac tttagttggg gttcgggttc    10620

gcgcgagatg atcacgtgcc ctgatttggt gtcgtccccc gtcgcgctgc gcacgtgatt    10680

tatttatttc cggtggctgc tgtctacgcg gggccttctc tgcccttctg tttcaacctt    10740

cgggcggttc tcgtaaccag cagtagcaat ccatttcgaa actcaaagag ctaaaaacgt    10800

taaacctcag cagtcgctcg acgaatgggc tgcggttggg aagcccacga ggcctatagc    10860

cagagcctcg agttgacagg agcccagacg ccttttccaa cggcaacttt tatataaaat    10920

ggcaatgtat tcatgcaatt gcggccgtgt caggttggag acactggacc acactctcca    10980

ttgcttcctg aggagatgga tcattgctag tgcatctacg cgcagcaatc ccgcaagctc    11040

gacaaccgta gatgggcttt ggtgggccaa tcaattacgc aacccgcacg ttaaattgta    11100

tgaggaagga aggccacggt acaaagtggg tggtcttcac ccagtggttg ttggtggcgt    11160

catgcagacc atggccgcca gtgtgctgga attgaatatt taccgttcgt ataatgtatg    11220

ctatacgaag ttataccggt ctcgtagtgt tcacgttcag ttcacggtga gcttaaaact    11280

atcttcaaga agagatttga gacctgattt atacttgcag caatgtttac ttcttatcgc    11340

gatacacgaa tgtgatacgg atcaaagtaa gcaggactac gataagataa cgaatgcggt    11400

gcagtccatg tcgattaggt atagatacat ttattttgtg ttatgttaca ttttgggggg    11460

atactgtcct acttgtagta cctacttgta gtggcgcgtt aggggcaggg catgctcatg    11520

tagagcgcct gccgctcgcc gtccgaggcg gtgccgtcgt acagggcggt gtccaggccg    11580

cagagggtga accccatccg ccggta                                         11606


<210>  148
<211>  5444
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Cas9, including a C-terminal SV40 nuclear 
       localization signal, codon optimized for expression in Yarrowia 
       lipolytica

<400>  148
gtgctactct actgatagca atagatgcgt cataattggt tggcccgctg agcctccaca       60

ggatactatt gcacataccc tggtcatgtg cagatcagct catttgtgga gactctggag      120

taacttagac gacgcctggt tcaattgccg caatgtgcgc ccacgcagat aatgtattga      180

ggggtggagc gcctcttggg gacttgctgt acttgtacgg gatattaaac gcactcagca      240

agaccatgac gtaaaacaca cctactgtac gatacgtact gtaggtattg tactcgtacc      300

cggtactaca aatagtacga tactatacgg agtgtatttg taccttgata tacgactggc      360

ggagtgaaga gaaggagttg aacaagacca gatggggata tcagccccag tgctttgtat      420

tacaagtacg agtacttaat agatactgta aggctattga tacggatggc agtaagtcat      480

tgagtaagca attgtggccc agcatctccc ctacgtactt gtaccatacc ccatggagac      540

accaatggtc tttcacgcac actgtcgtgt gctgtatcgc agaatcgggt gtccaaccaa      600

atgccgttac ccccacgtca cagccgatag acagatacac catcaatacc agcaggttgt      660

atcatgcggt tggctgaagg taagctgatt ggtctaaaaa ctgtagctgt cctaattcaa      720

cgagcgctat ttggggccaa ccacctcggc caagcggcct ttaatctgcg tgccccagag      780

gcgtctaatg aggctctggc cgccactgta ggagtgtttc tctgtgcgca cacgcagttt      840

tgagtttggg cgactttccc tttttcccaa ttgcgtacac acacagctcc gagctaagcg      900

ctgtccttga accttctccc tcttttccct ctttttctct tccccttccc ctcctccaca      960

ttaaggccaa atcctgaatt gcaccaacta gtacaacgac aacaatggac aagaagtact     1020

ccatcggttt ggacattggt actaactctg tcggctgggc cgtcatcacc gacgagtaca     1080

aggttccctc caagaagttc aaggtccttg gcaacaccga ccgacactct atcaagaaga     1140

acctgatcgg tgctctgctg ttcgactctg gcgagactgc cgaggccacc cgactgaagc     1200

gaaccgctcg acgccgatac acccgacgaa agaaccgaat ctgttacctc caggagatct     1260

tcagcaacga gatggctaag gtcgacgact ccttcttcca ccgactcgag gagtctttcc     1320

tggtcgaaga ggataagaag cacgagcgac accccatctt cggcaacatt gttgatgagg     1380

ttgcctacca tgagaagtac cccaccatct accacctccg aaagaagctc gtcgactcca     1440

ctgacaaggc tgacctccga ctcatctacc ttgctctcgc ccacatgatc aagttccgag     1500

gtcacttcct cattgagggt gatctcaacc ccgacaactc cgacgttgac aagctgttca     1560

tccagctcgt ccagacctac aaccagctct ttgaggagaa ccctatcaac gcttctggtg     1620

ttgacgccaa ggccattctc tccgcccgac tctctaagtc ccgacgactc gagaacctca     1680

ttgcccagct gcccggcgag aagaagaacg gcctcttcgg taacctgatt gctctctctc     1740

ttggtctgac ccccaacttc aagtccaact ttgacctcgc cgaggacgcc aagctccagc     1800

tgtccaagga cacctacgat gacgatctgg acaacctcct ggcccagatc ggtgaccagt     1860

acgccgatct cttccttgcc gccaagaacc tctccgacgc catcctgctc tccgacatcc     1920

tccgagtcaa caccgagatt accaaggctc ctctgtctgc ctctatgatc aagcgatacg     1980

acgagcacca ccaggatctc actcttctca aggctctcgt ccgacagcag ctccccgaga     2040

agtacaagga gattttcttt gaccagtcca agaacggtta cgctggctac attgacggtg     2100

gtgcttccca ggaagagttt tacaagttca tcaagcctat tctggagaag atggacggta     2160

ccgaggagct gctcgtcaag ctcaaccgag aggacctcct tcgaaagcag cgaaccttcg     2220

ataacggctc catcccccac cagatccacc tgggtgagct ccacgccatt ctccgaagac     2280

aagaggactt ctaccccttc ctaaaggata accgagagaa gatcgagaag attctcacct     2340

tccgaatccc ctactacgtc ggtcccctcg ctcgaggtaa ctcccgattt gcttggatga     2400

cccgaaagtc cgaggagact atcaccccct ggaactttga agaggtagtc gacaagggtg     2460

cctccgccca gtctttcatt gagcggatga ccaacttcga taagaacctc cccaacgaga     2520

aggtccttcc caagcactct ctcctctacg agtacttcac cgtctacaac gagctgacca     2580

aggtcaagta cgttaccgag ggcatgcgaa agcccgcttt cctctctggt gagcagaaga     2640

aggccattgt cgacctcctg ttcaagacta accgaaaagt caccgtcaag cagctcaagg     2700

aagactactt caagaagatt gagtgcttcg actccgtcga gatttccggt gtcgaggacc     2760

gattcaacgc ctccctcggc acctaccacg atcttctgaa gatcatcaag gacaaggact     2820

ttcttgataa cgaggagaac gaggacattc tcgaggacat cgtcctcacc ctcacccttt     2880

tcgaggatcg agagatgatc gaggagcgac tcaagaccta cgcccatctc ttcgacgaca     2940

aggtcatgaa gcaactcaag cgacgacgat acactggctg gggccgactt tcccgaaagc     3000

tcatcaacgg catccgagac aagcagtctg gcaagaccat cctggacttc ctgaagtccg     3060

acggtttcgc caaccgaaac ttcatgcagc tcatccacga cgactctctt accttcaaag     3120

aggatatcca gaaggcccag gtttctggcc agggcgactc cctccacgag cacattgcca     3180

acctcgccgg atcccccgcc atcaaaaagg gtatcctcca gaccgtcaag gttgtcgacg     3240

aactcgtgaa ggtcatgggc cgacacaagc ccgagaacat cgttatcgag atggcccgag     3300

agaaccagac cacccagaag ggtcagaaga actcccgaga gcgaatgaag cgaatcgaag     3360

agggtatcaa ggagctcggt tcccagattc tcaaggagca ccccgtcgag aacacccagc     3420

tccagaacga gaaactctac ctgtactacc tccagaatgg ccgagacatg tacgttgacc     3480

aggagctcga catcaaccga ctctccgact acgacgtcga ccacattgtt cctcagtcct     3540

tcctcaagga cgactccatc gacaacaagg ttctgacccg atctgacaag aaccgaggta     3600

agtccgacaa cgttccctcc gaagaggtcg ttaagaagat gaagaactac tggcgacagc     3660

ttctcaacgc caaactgatc acccagcgaa agtttgacaa cctcaccaag gccgagcgag     3720

gtggtctgtc cgagctggac aaggccggct tcattaagcg acagctggtc gagactcgac     3780

agatcaccaa gcacgtcgcc cagatcctcg actcccgaat gaacaccaag tacgacgaga     3840

acgacaagct catccgggag gtcaaggtca tcaccctgaa gtctaagctt gtctccgact     3900

tccgaaagga cttccagttc tacaaggtcc gagagatcaa caactaccac cacgcccacg     3960

acgcctacct caacgccgtt gttggtaccg ccctcatcaa gaagtatccc aagctcgagt     4020

ccgagttcgt ttacggcgac tacaaggttt acgatgtccg aaagatgatt gccaagtccg     4080

agcaggagat cggtaaggcc accgccaagt actttttcta ctccaacatc atgaatttct     4140

tcaagaccga gatcactctc gccaacggtg agattcgaaa gcgacccctg attgagacta     4200

atggtgagac tggtgagatc gtctgggata agggccgaga cttcgccacc gtccgaaagg     4260

tcctgtccat gccccaggtc aacattgtca agaagaccga ggtccagacc ggtggcttct     4320

ccaaggagtc cattctcccc aagcgaaact ccgacaaact catcgcccgt aagaaggact     4380

gggatccgaa gaagtacggt ggtttcgatt ctcccaccgt tgcctactcc gtcctcgttg     4440

ttgctaaagt cgagaagggt aagtctaaga aactcaagtc cgtgaaggag ctactcggta     4500

tcaccatcat ggagcgatct tcttttgaga agaaccccat tgacttcctc gaggccaagg     4560

gttacaaaga ggtcaagaag gacctgatta tcaagctgcc caagtactcc ctctttgagc     4620

tcgagaacgg ccgaaagcga atgctggctt ccgctggtga gctgcagaag ggcaacgagc     4680

tcgctctgcc ctccaagtac gtcaacttcc tctacctggc ctcccactac gagaagctca     4740

agggctcccc cgaggacaac gagcagaagc agctgttcgt tgagcagcac aagcactacc     4800

tcgacgagat catcgagcag atctccgagt tctccaagcg agtcatcctc gctgacgcca     4860

accttgataa ggttctctct gcttacaaca agcaccggga caagcccatc cgagagcagg     4920

ccgagaatat catccacctc ttcactctca ccaacctcgg cgctcctgct gccttcaagt     4980

acttcgacac caccattgac cgaaagaggt acacctccac caaggaagtc ctcgacgcca     5040

ccctgatcca ccagtccatc accggcctct acgaaacccg aatcgacctc tcccagctcg     5100

gcggtgactc tcgagccgac cccaagaaga agcgaaaagt ctaaatatcc gaagatcaag     5160

agcgaagcaa gttgtaagtc caggacatgt ttcccgccca cgcgagtgat ttataacacc     5220

tctctttttt gacacccgct cgccttgaaa ttcatgtcac ataaattata gtcaacgacg     5280

tttgaataac ttgtcttgta gttcgatgat gatcatatga ttacattaat agtaattact     5340

gtatttgata tatatactaa ttacaatagt acatattaga acatacaata gttagtgccg     5400

tgaagtggct taaaataccg cgagtcgatt acgtaatatt atta                      5444


<210>  149
<211>  1004
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Yarrowia Yl_007 promoter

<400>  149
gtgctactct actgatagca atagatgcgt cataattggt tggcccgctg agcctccaca       60

ggatactatt gcacataccc tggtcatgtg cagatcagct catttgtgga gactctggag      120

taacttagac gacgcctggt tcaattgccg caatgtgcgc ccacgcagat aatgtattga      180

ggggtggagc gcctcttggg gacttgctgt acttgtacgg gatattaaac gcactcagca      240

agaccatgac gtaaaacaca cctactgtac gatacgtact gtaggtattg tactcgtacc      300

cggtactaca aatagtacga tactatacgg agtgtatttg taccttgata tacgactggc      360

ggagtgaaga gaaggagttg aacaagacca gatggggata tcagccccag tgctttgtat      420

tacaagtacg agtacttaat agatactgta aggctattga tacggatggc agtaagtcat      480

tgagtaagca attgtggccc agcatctccc ctacgtactt gtaccatacc ccatggagac      540

accaatggtc tttcacgcac actgtcgtgt gctgtatcgc agaatcgggt gtccaaccaa      600

atgccgttac ccccacgtca cagccgatag acagatacac catcaatacc agcaggttgt      660

atcatgcggt tggctgaagg taagctgatt ggtctaaaaa ctgtagctgt cctaattcaa      720

cgagcgctat ttggggccaa ccacctcggc caagcggcct ttaatctgcg tgccccagag      780

gcgtctaatg aggctctggc cgccactgta ggagtgtttc tctgtgcgca cacgcagttt      840

tgagtttggg cgactttccc tttttcccaa ttgcgtacac acacagctcc gagctaagcg      900

ctgtccttga accttctccc tcttttccct ctttttctct tccccttccc ctcctccaca      960

ttaaggccaa atcctgaatt gcaccaacta gtacaacgac aaca                      1004


<210>  150
<211>  300
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Yarrowia Yl_GPD terminator

<400>  150
atatccgaag atcaagagcg aagcaagttg taagtccagg acatgtttcc cgcccacgcg       60

agtgatttat aacacctctc ttttttgaca cccgctcgcc ttgaaattca tgtcacataa      120

attatagtca acgacgtttg aataacttgt cttgtagttc gatgatgatc atatgattac      180

attaatagta attactgtat ttgatatata tactaattac aatagtacat attagaacat      240

acaatagtta gtgccgtgaa gtggcttaaa ataccgcgag tcgattacgt aatattatta      300


<210>  151
<211>  12810
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of pSTV089

<400>  151
ggttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta       60

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag      120

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg      180

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg      240

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg      300

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga      360

agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc      420

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt      480

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact      540

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg      600

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt      660

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt      720

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct      780

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg      840

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt      900

aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt      960

gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc     1020

gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg     1080

cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc     1140

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg     1200

gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca     1260

ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga     1320

tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct     1380

ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg     1440

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca     1500

accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca     1560

cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct     1620

tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact     1680

cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa     1740

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc     1800

atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga     1860

tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga     1920

aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg     1980

cgtatcacga ggccctttcg tctggcctag gaagcgactt ccaatcgctt tgcatatcca     2040

gtaccacacc cacaggcgtt tgtgctactc tactgatagc aatagatgcg tcataattgg     2100

ttggcccgct gagcctccac aggatactat tgcacatacc ctggtcatgt gcagatcagc     2160

tcatttgtgg agactctgga gtaacttaga cgacgcctgg ttcaattgcc gcaatgtgcg     2220

cccacgcaga taatgtattg aggggtggag cgcctcttgg ggacttgctg tacttgtacg     2280

ggatattaaa cgcactcagc aagaccatga cgtaaaacac acctactgta cgatacgtac     2340

tgtaggtatt gtactcgtac ccggtactac aaatagtacg atactatacg gagtgtattt     2400

gtaccttgat atacgactgg cggagtgaag agaaggagtt gaacaagacc agatggggat     2460

atcagcccca gtgctttgta ttacaagtac gagtacttaa tagatactgt aaggctattg     2520

atacggatgg cagtaagtca ttgagtaagc aattgtggcc cagcatctcc cctacgtact     2580

tgtaccatac cccatggaga caccaatggt ctttcacgca cactgtcgtg tgctgtatcg     2640

cagaatcggg tgtccaacca aatgccgtta cccccacgtc acagccgata gacagataca     2700

ccatcaatac cagcaggttg tatcatgcgg ttggctgaag gtaagctgat tggtctaaaa     2760

actgtagctg tcctaattca acgagcgcta tttggggcca accacctcgg ccaagcggcc     2820

tttaatctgc gtgccccaga ggcgtctaat gaggctctgg ccgccactgt aggagtgttt     2880

ctctgtgcgc acacgcagtt ttgagtttgg gcgactttcc ctttttccca attgcgtaca     2940

cacacagctc cgagctaagc gctgtccttg aaccttctcc ctcttttccc tctttttctc     3000

ttccccttcc cctcctccac attaaggcca aatcctgaat tgcaccaact agtacaacga     3060

caacaatgga caagaagtac tccatcggtt tggacattgg tactaactct gtcggctggg     3120

ccgtcatcac cgacgagtac aaggttccct ccaagaagtt caaggtcctt ggcaacaccg     3180

accgacactc tatcaagaag aacctgatcg gtgctctgct gttcgactct ggcgagactg     3240

ccgaggccac ccgactgaag cgaaccgctc gacgccgata cacccgacga aagaaccgaa     3300

tctgttacct ccaggagatc ttcagcaacg agatggctaa ggtcgacgac tccttcttcc     3360

accgactcga ggagtctttc ctggtcgaag aggataagaa gcacgagcga caccccatct     3420

tcggcaacat tgttgatgag gttgcctacc atgagaagta ccccaccatc taccacctcc     3480

gaaagaagct cgtcgactcc actgacaagg ctgacctccg actcatctac cttgctctcg     3540

cccacatgat caagttccga ggtcacttcc tcattgaggg tgatctcaac cccgacaact     3600

ccgacgttga caagctgttc atccagctcg tccagaccta caaccagctc tttgaggaga     3660

accctatcaa cgcttctggt gttgacgcca aggccattct ctccgcccga ctctctaagt     3720

cccgacgact cgagaacctc attgcccagc tgcccggcga gaagaagaac ggcctcttcg     3780

gtaacctgat tgctctctct cttggtctga cccccaactt caagtccaac tttgacctcg     3840

ccgaggacgc caagctccag ctgtccaagg acacctacga tgacgatctg gacaacctcc     3900

tggcccagat cggtgaccag tacgccgatc tcttccttgc cgccaagaac ctctccgacg     3960

ccatcctgct ctccgacatc ctccgagtca acaccgagat taccaaggct cctctgtctg     4020

cctctatgat caagcgatac gacgagcacc accaggatct cactcttctc aaggctctcg     4080

tccgacagca gctccccgag aagtacaagg agattttctt tgaccagtcc aagaacggtt     4140

acgctggcta cattgacggt ggtgcttccc aggaagagtt ttacaagttc atcaagccta     4200

ttctggagaa gatggacggt accgaggagc tgctcgtcaa gctcaaccga gaggacctcc     4260

ttcgaaagca gcgaaccttc gataacggct ccatccccca ccagatccac ctgggtgagc     4320

tccacgccat tctccgaaga caagaggact tctacccctt cctaaaggat aaccgagaga     4380

agatcgagaa gattctcacc ttccgaatcc cctactacgt cggtcccctc gctcgaggta     4440

actcccgatt tgcttggatg acccgaaagt ccgaggagac tatcaccccc tggaactttg     4500

aagaggtagt cgacaagggt gcctccgccc agtctttcat tgagcggatg accaacttcg     4560

ataagaacct ccccaacgag aaggtccttc ccaagcactc tctcctctac gagtacttca     4620

ccgtctacaa cgagctgacc aaggtcaagt acgttaccga gggcatgcga aagcccgctt     4680

tcctctctgg tgagcagaag aaggccattg tcgacctcct gttcaagact aaccgaaaag     4740

tcaccgtcaa gcagctcaag gaagactact tcaagaagat tgagtgcttc gactccgtcg     4800

agatttccgg tgtcgaggac cgattcaacg cctccctcgg cacctaccac gatcttctga     4860

agatcatcaa ggacaaggac tttcttgata acgaggagaa cgaggacatt ctcgaggaca     4920

tcgtcctcac cctcaccctt ttcgaggatc gagagatgat cgaggagcga ctcaagacct     4980

acgcccatct cttcgacgac aaggtcatga agcaactcaa gcgacgacga tacactggct     5040

ggggccgact ttcccgaaag ctcatcaacg gcatccgaga caagcagtct ggcaagacca     5100

tcctggactt cctgaagtcc gacggtttcg ccaaccgaaa cttcatgcag ctcatccacg     5160

acgactctct taccttcaaa gaggatatcc agaaggccca ggtttctggc cagggcgact     5220

ccctccacga gcacattgcc aacctcgccg gatcccccgc catcaaaaag ggtatcctcc     5280

agaccgtcaa ggttgtcgac gaactcgtga aggtcatggg ccgacacaag cccgagaaca     5340

tcgttatcga gatggcccga gagaaccaga ccacccagaa gggtcagaag aactcccgag     5400

agcgaatgaa gcgaatcgaa gagggtatca aggagctcgg ttcccagatt ctcaaggagc     5460

accccgtcga gaacacccag ctccagaacg agaaactcta cctgtactac ctccagaatg     5520

gccgagacat gtacgttgac caggagctcg acatcaaccg actctccgac tacgacgtcg     5580

accacattgt tcctcagtcc ttcctcaagg acgactccat cgacaacaag gttctgaccc     5640

gatctgacaa gaaccgaggt aagtccgaca acgttccctc cgaagaggtc gttaagaaga     5700

tgaagaacta ctggcgacag cttctcaacg ccaaactgat cacccagcga aagtttgaca     5760

acctcaccaa ggccgagcga ggtggtctgt ccgagctgga caaggccggc ttcattaagc     5820

gacagctggt cgagactcga cagatcacca agcacgtcgc ccagatcctc gactcccgaa     5880

tgaacaccaa gtacgacgag aacgacaagc tcatccggga ggtcaaggtc atcaccctga     5940

agtctaagct tgtctccgac ttccgaaagg acttccagtt ctacaaggtc cgagagatca     6000

acaactacca ccacgcccac gacgcctacc tcaacgccgt tgttggtacc gccctcatca     6060

agaagtatcc caagctcgag tccgagttcg tttacggcga ctacaaggtt tacgatgtcc     6120

gaaagatgat tgccaagtcc gagcaggaga tcggtaaggc caccgccaag tactttttct     6180

actccaacat catgaatttc ttcaagaccg agatcactct cgccaacggt gagattcgaa     6240

agcgacccct gattgagact aatggtgaga ctggtgagat cgtctgggat aagggccgag     6300

acttcgccac cgtccgaaag gtcctgtcca tgccccaggt caacattgtc aagaagaccg     6360

aggtccagac cggtggcttc tccaaggagt ccattctccc caagcgaaac tccgacaaac     6420

tcatcgcccg taagaaggac tgggatccga agaagtacgg tggtttcgat tctcccaccg     6480

ttgcctactc cgtcctcgtt gttgctaaag tcgagaaggg taagtctaag aaactcaagt     6540

ccgtgaagga gctactcggt atcaccatca tggagcgatc ttcttttgag aagaacccca     6600

ttgacttcct cgaggccaag ggttacaaag aggtcaagaa ggacctgatt atcaagctgc     6660

ccaagtactc cctctttgag ctcgagaacg gccgaaagcg aatgctggct tccgctggtg     6720

agctgcagaa gggcaacgag ctcgctctgc cctccaagta cgtcaacttc ctctacctgg     6780

cctcccacta cgagaagctc aagggctccc ccgaggacaa cgagcagaag cagctgttcg     6840

ttgagcagca caagcactac ctcgacgaga tcatcgagca gatctccgag ttctccaagc     6900

gagtcatcct cgctgacgcc aaccttgata aggttctctc tgcttacaac aagcaccggg     6960

acaagcccat ccgagagcag gccgagaata tcatccacct cttcactctc accaacctcg     7020

gcgctcctgc tgccttcaag tacttcgaca ccaccattga ccgaaagagg tacacctcca     7080

ccaaggaagt cctcgacgcc accctgatcc accagtccat caccggcctc tacgaaaccc     7140

gaatcgacct ctcccagctc ggcggtgact ctcgagccga ccccaagaag aagcgaaaag     7200

tctaaatatc cgaagatcaa gagcgaagca agttgtaagt ccaggacatg tttcccgccc     7260

acgcgagtga tttataacac ctctcttttt tgacacccgc tcgccttgaa attcatgtca     7320

cataaattat agtcaacgac gtttgaataa cttgtcttgt agttcgatga tgatcatatg     7380

attacattaa tagtaattac tgtatttgat atatatacta attacaatag tacatattag     7440

aacatacaat agttagtgcc gtgaagtggc ttaaaatacc gcgagtcgat tacgtaatat     7500

tattacctct tgcccatcga acgtacaagt actcctctgt tctctccttc ctttgctttg     7560

tgcacgaaga actgcggtca ggtgacacaa ctttttccat ctcagggtgt gtcgcgtgtg     7620

cttcatccaa actttagttg gggttcgggt tcgcgcgaga tgatcacgtg ccctgatttg     7680

gtgtcgtccc ccgtcgcgct gcgcacgtga tttatttatt tccggtggct gctgtctacg     7740

cggggccttc tctgcccttc tgtttcaacc ttcgggcggt tctcgtaacc agcagtagca     7800

atccatttcg aaactcaaag agctaaaaac gttaaacctc agcagtcgct cgacgaatgg     7860

gctgcggttg ggaagcccac gaggcctata gccagagcct cgagttgaca ggagcccaga     7920

cgccttttcc aacggcaact tttatataaa atggcaatgt attcatgcaa ttgcggccgt     7980

gtcaggttgg agacactgga ccacactctc cattgcttcc tgaggagatg gatcattgct     8040

agtgcatcta cgcgcagcaa tcccgcaagc tcgacaaccg tagatgggct ttggtgggcc     8100

aatcaattac gcaacccgca cgttaaattg tatgaggaag gaaggccacg gtacaaagtg     8160

ggtggtcttc acccagtggt tgttggtggc gtcatgcaga ccatgcattg gggatagcac     8220

agggttgggg tgtcttgtgg actcaatggg tgaaaggaga tggaaaaggg cggtgaaaag     8280

tggtagaatc gaaatccctg acgtcaattt ataaagtaaa atgcgtttct gccattttgc     8340

tcccctcctt ctttcgcaat cgcctcccca aaagttgtcg tggcagtaca catgcttgca     8400

tacaatgaag ctaatccggc ttgctcagta gttgctatat ccaggcatgg tgtgaaaccc     8460

ctcaaagtat atataggagc ggtgagcccc agtctggggt cttttctctc catctcaaaa     8520

ctactttctc acaatgcgat atctgatgag tccgtgagga cgaaacgagt aagctcgtca     8580

tatcgccgca agattacacg ttttagagct agaaatagca agttaaaata aggctagtcc     8640

gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttg gccggcatgg tcccagcctc     8700

ctcgctggcg ccggctgggc aacatgcttc ggcatggcga atgggactaa acttcgagct     8760

aatccagtag cttacgttac ccaggggcag gtcaactggc tagccacgag tctgtcccag     8820

gtcgcaattt agtgtaataa acaatatata tattgagtct aaagggaatt gtagctattg     8880

tgattgtgtg attttcgtct tgctggttct tattgtgtcc cattcgtttc atcctgatga     8940

ggacccctgg aaccggtgtt ttcttagtct ctgcaatcgc tagtcttgtt gctatgacag     9000

ttgcgtcgac actattcagg tcatctatcg gttattctga tattataata cctccggatc     9060

gatgtacctg atttatactt gcagcaatgt ttacttctta tcgcgataca cgaatgtgat     9120

acggatcaaa gtaagcagga ctacgataag ataacgaatg cggtgcagtc catgtcgatt     9180

aggtatagat acatttattt tgtgttatgt tacattttgg ggggatactg tcctacttgt     9240

agtacctact tgtagtggcg cgtctattcc tttgccctcg gacgagtgct ggggcgtcgg     9300

tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg     9360

gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc     9420

tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga     9480

ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc     9540

tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg     9600

gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg     9660

cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact     9720

tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg     9780

acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg     9840

aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc     9900

gtctggctaa gatcggccgc agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca     9960

gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg    10020

caataggtca ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg    10080

gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt    10140

acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc    10200

tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca    10260

gacgtcgcgg tgagttcagg ctttttcata tgggtacctg agaacatttt tgtgtctagg    10320

tgtttgtgtt tggactgcga tcagtgaaga aaagaagagg aaaaattgtg caagaaattt    10380

tgctttcaag acttggctga tgcagcaggg taactctggg acacagacct atgtttgtgg    10440

ttaaactcaa tgcacgtggt acgtgcgtgg agcgcttacc catccaaggg tgtggacatg    10500

gaaccgacgg tccgtggagt tgtgtaatgt cattttggcg actcttgaag caaggctata    10560

aaaaaattgt gtggcttgag tcttatcgag ctcggtcact acaagagtta atcttcctgt    10620

ctcaggcaga caggtcaggc agggttactt ttgggtgtgc tgtaactcac tgtatggccg    10680

ttagtgcgca tagacgttgt acatactgga ccgaattgta gcgtgctcaa tagggccaat    10740

aaagctattg tagggatccg aattttcaga acctaattta tctgttaccc ggcctgtggc    10800

tcgcacagct taaaaatggt caaactttcc ccttcttgtc tttttttcct cacattcatc    10860

aggttcttgt cttgatcttt caagtgagta ttaattaccg accttggttc ttcattggga    10920

gagcattgga agccgtggtg cagcaaccac aaaacggttc ttccccttcg ataccttctt    10980

gcctgccttt caatacaagt cggctcgatt agcggtggtc gcccccgcca gcggagaaca    11040

tggaactaac ccagaatgag agctaagtgg agaaagaaga gagtcagacg actcaagcga    11100

aagcgccgca aggtccgagc tcgatccaaa taagcggttt ttaacggaga tttaacacta    11160

aatcgaagaa cttttcccgt ttcatttgcg aatgagctcg ttaacaaaat cccccagttt    11220

ttttatccag ctgtaaggat tgacattagt aatgaattat tgtttggtat atttaaatct    11280

gtagttcctt tctgtccgtg tcggcaactg tcgtactcgt gatttacttg tattgacgaa    11340

tacttactgt agcgcactct gctgctactg gtcgtaagga tgtgctattt cggtgtatgg    11400

tgggtttttt gggggtcgga accgaagact gttacacggg cacggctcgt tgtgtacacg    11460

cacagagctc ttgcgagtca tgttgtagct agctcgtcgt gttcaggaac tgttcgatgg    11520

ttcggagaga gtcgccgccc agaacatacg cgcaccgatg tcagcagaca gccttattac    11580

aagtatattc aagcaagtat atccgtaggg tgcgggtgat ttggatctaa ggttcgtact    11640

caacactcac gagcagcttg cctatgttac atccttttat cagacataac ataattggag    11700

tttacttaca cacggggtgt acctgtatga gcaccaccta caattgtagc actggtactt    11760

gtacaaagaa tttattcgta cgaatcacag ggacggccgc cctcaccgaa ccagcgaata    11820

cctcagcggt cccctgcagt gactcaacaa agcgatatga acatcttgcg atggtatcct    11880

gctgatagtt tttactgtac aaacacctgt gtagctcctt ctagcatttt taagttattc    11940

acacctcaag gggagggata aattaaataa attccaaaag cgaagatcga gaaactaaat    12000

taaaattcca aaaacgaagt tggaacacaa ccccccgaaa aaaaacaaca aacaaaaaac    12060

ccaacaaaat aaacaaaaac aaaataaata tataactacc agtatctgac taaaagttca    12120

aatactcgta cttacaacaa atagaaatga gccggccaaa attctgcaga aaaaaatttc    12180

aaacaagtac tggtataatt aaattaaaaa acacatcaaa gtatcataac gttagttatt    12240

ttattttatt taataaaaga aaacaacaag atgggctcaa aactttcaac ttatacgata    12300

cataccaaat aacaatttag tatttatcta agtgcttttc gtagataatg gaatacaaat    12360

ggatatccag agtatacaca tggatagtat acactgacac gacaattctg tatctcttta    12420

tgttaactac tgtgaggcat taaatagagc ttgatatata aaatgttaca tttcacagtc    12480

tgaacttttg cagattacct aatttggtaa gatattaatt atgaactgaa agttgatggc    12540

atccctaaat ttgatgaaag atgaaattgt aaatgaggtg gtaaaagagc tacagtcgtt    12600

ttgttttgag ataccatcat ctctaacgaa atatctatta aaaatctcag tgtgatcatg    12660

agtcattgcc atcctggaaa atgtcatcat ggctgatatt tctaactgtt tacttgagat    12720

aaatatatat ttacaagaac ttcccttgaa attaatttag atataaaatg tttgcgggca    12780

agttactacg aggaataaat tatatctaga                                     12810


<210>  152
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20-bp genomic target of the KU70 gene

<400>  152
atatcgccgc aagattacac                                                   20


<210>  153
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 100-bp donor DNA fragment used for 
       knocking out the KU70 gene in the Yarrowia genome

<400>  153
catcgcgaca cgaacacgaa acacgaacca cgaaccgccg ctttttgaaa ctagggaggc       60

acatctaaac gaataacgaa tattaatgat accatcatat                            100


<210>  154
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to confirm knock out of
       KU70 gene in the Yarrowia genome

<400>  154
gagcacgtca cgtgttctcc                                                   20


<210>  155
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to confirm knock out of
       KU70 gene in the Yarrowia genome

<400>  155
agtggtaccg ctactttgtg                                                   20


<210>  156
<211>  2042
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the GFP expression cassette (Yl_HSP.pro - 
       A.vic_eGFP ORF - Yl_GPD.ter)

<400>  156
gtgcaatcac atgttgctac tgtacctgct gtggaccacg cacggcggaa cgtaccgtac       60

aaatattttc ttgctcacat gactctctct cggccgcgca cgccggtggc aaattgctct      120

tgcattggct ctgtctctag acgtccaaac cgtccaaagt ggcagggtga cgtgatgcga      180

cgcacgaagg agatggcccg gtggcgagga accggacacg gcgagccggc gggaaaaaag      240

gcggaaaacg aaaagcgaag ggcacaatct gacggtgcgg ctgccaccaa cccaaggagg      300

ctattttggg tcgctttcca tttcacattc gccctcaatg gccactttgc ggtggtgaac      360

atggtttctg aaacaacccc ccagaattag agtatattga tgtgtttaag attgggttgc      420

tatttggcca ttgtggggga gggtagcgac gtggaggaca ttccagggcg aattgagcct      480

agaaagtggt agcattccaa ccgtctaagt cgtccgaatt gatcgctata actatcacct      540

ctctcacatg tctacttccc caaccaacat ccccaacctc ccccacacta aagttcacgc      600

caataatgta ggcactcttt ctgggtgtgg gacagcagag caatacggag gggagattac      660

acaacgagcc acaattgggg agatggtagc catctcactc gacccgtcga cttttggcaa      720

cgctcaatta cccaccaaat ttgggctgga gttgagggga ccgtgttcca gcgctgtagg      780

accagcaaca cacacggtat caacagcaac caacgccccc gctaatgcac ccagtactgc      840

gcaggtgtgg gccaggtgcg ttccagatgc gagttggcga accctaagcc gacagtgtac      900

tttttgggac gggcagtagc aatcgtgggc ggaaaccccg gtgtatataa aggggtggag      960

aggacggatt attagcacca acacacacac ttatactaca atggagggtg atattcacgg     1020

tgcttctaag ggtgaggagc ttttcaccgg cgttgtccct attctcgttg agcttgatgg     1080

tgatgtcaac ggtcacaagt tttctgtctc tggtgagggt gagggtgatg ctacttacgg     1140

taagctcacc ctcaagttta tttgtaccac cggcaagctc cccgttccct ggcctaccct     1200

cgttaccact ctcacctacg gtgttcagtg tttttcccga taccccgatc acatgaagcg     1260

acacgacttt ttcaagtctg ccatgcccga gggttacgtt caggagcgaa ctatttcttt     1320

caaggatgac ggtaactaca agacccgagc tgaggttaag tttgagggtg ataccctcgt     1380

taaccgaatc gagcttaagg gtattgattt caaggaggat ggtaacattc tcggtcacaa     1440

gctggagtac aactacaact ctcacaacgt ttacatcacc gctgacaagc agaagaacgg     1500

tatcaaggct aacttcaaga ttcgacacaa cattgaggat ggttccgttc agcttgctga     1560

ccactaccag cagaacactc ccattggcga tggccctgtc ctcctccccg acaaccacta     1620

cctctctacc cagtctgccc tttctaagga ccccaacgag aagcgagatc acatggtcct     1680

ccttgagttc gttaccgctg ctggtattac tcacggtatg gatgagctct acaagtaaat     1740

aaatatccga agatcaagag cgaagcaagt tgtaagtcca ggacatgttt cccgcccacg     1800

cgagtgattt ataacacctc tcttttttga cacccgctcg ccttgaaatt catgtcacat     1860

aaattatagt caacgacgtt tgaataactt gtcttgtagt tcgatgatga tcatatgatt     1920

acattaatag taattactgt atttgatata tatactaatt acaatagtac atattagaac     1980

atacaatagt tagtgccgtg aagtggctta aaataccgcg agtcgattac gtaatattat     2040

ta                                                                    2042


<210>  157
<211>  12810
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of plasmid pSTV086

<400>  157
ggttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta       60

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag      120

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg      180

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg      240

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg      300

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga      360

agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc      420

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt      480

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact      540

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg      600

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt      660

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt      720

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct      780

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg      840

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt      900

aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt      960

gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc     1020

gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg     1080

cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc     1140

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg     1200

gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca     1260

ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga     1320

tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct     1380

ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg     1440

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca     1500

accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca     1560

cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct     1620

tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact     1680

cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa     1740

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc     1800

atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga     1860

tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga     1920

aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg     1980

cgtatcacga ggccctttcg tctggcctag gaagcgactt ccaatcgctt tgcatatcca     2040

gtaccacacc cacaggcgtt tgtgctactc tactgatagc aatagatgcg tcataattgg     2100

ttggcccgct gagcctccac aggatactat tgcacatacc ctggtcatgt gcagatcagc     2160

tcatttgtgg agactctgga gtaacttaga cgacgcctgg ttcaattgcc gcaatgtgcg     2220

cccacgcaga taatgtattg aggggtggag cgcctcttgg ggacttgctg tacttgtacg     2280

ggatattaaa cgcactcagc aagaccatga cgtaaaacac acctactgta cgatacgtac     2340

tgtaggtatt gtactcgtac ccggtactac aaatagtacg atactatacg gagtgtattt     2400

gtaccttgat atacgactgg cggagtgaag agaaggagtt gaacaagacc agatggggat     2460

atcagcccca gtgctttgta ttacaagtac gagtacttaa tagatactgt aaggctattg     2520

atacggatgg cagtaagtca ttgagtaagc aattgtggcc cagcatctcc cctacgtact     2580

tgtaccatac cccatggaga caccaatggt ctttcacgca cactgtcgtg tgctgtatcg     2640

cagaatcggg tgtccaacca aatgccgtta cccccacgtc acagccgata gacagataca     2700

ccatcaatac cagcaggttg tatcatgcgg ttggctgaag gtaagctgat tggtctaaaa     2760

actgtagctg tcctaattca acgagcgcta tttggggcca accacctcgg ccaagcggcc     2820

tttaatctgc gtgccccaga ggcgtctaat gaggctctgg ccgccactgt aggagtgttt     2880

ctctgtgcgc acacgcagtt ttgagtttgg gcgactttcc ctttttccca attgcgtaca     2940

cacacagctc cgagctaagc gctgtccttg aaccttctcc ctcttttccc tctttttctc     3000

ttccccttcc cctcctccac attaaggcca aatcctgaat tgcaccaact agtacaacga     3060

caacaatgga caagaagtac tccatcggtt tggacattgg tactaactct gtcggctggg     3120

ccgtcatcac cgacgagtac aaggttccct ccaagaagtt caaggtcctt ggcaacaccg     3180

accgacactc tatcaagaag aacctgatcg gtgctctgct gttcgactct ggcgagactg     3240

ccgaggccac ccgactgaag cgaaccgctc gacgccgata cacccgacga aagaaccgaa     3300

tctgttacct ccaggagatc ttcagcaacg agatggctaa ggtcgacgac tccttcttcc     3360

accgactcga ggagtctttc ctggtcgaag aggataagaa gcacgagcga caccccatct     3420

tcggcaacat tgttgatgag gttgcctacc atgagaagta ccccaccatc taccacctcc     3480

gaaagaagct cgtcgactcc actgacaagg ctgacctccg actcatctac cttgctctcg     3540

cccacatgat caagttccga ggtcacttcc tcattgaggg tgatctcaac cccgacaact     3600

ccgacgttga caagctgttc atccagctcg tccagaccta caaccagctc tttgaggaga     3660

accctatcaa cgcttctggt gttgacgcca aggccattct ctccgcccga ctctctaagt     3720

cccgacgact cgagaacctc attgcccagc tgcccggcga gaagaagaac ggcctcttcg     3780

gtaacctgat tgctctctct cttggtctga cccccaactt caagtccaac tttgacctcg     3840

ccgaggacgc caagctccag ctgtccaagg acacctacga tgacgatctg gacaacctcc     3900

tggcccagat cggtgaccag tacgccgatc tcttccttgc cgccaagaac ctctccgacg     3960

ccatcctgct ctccgacatc ctccgagtca acaccgagat taccaaggct cctctgtctg     4020

cctctatgat caagcgatac gacgagcacc accaggatct cactcttctc aaggctctcg     4080

tccgacagca gctccccgag aagtacaagg agattttctt tgaccagtcc aagaacggtt     4140

acgctggcta cattgacggt ggtgcttccc aggaagagtt ttacaagttc atcaagccta     4200

ttctggagaa gatggacggt accgaggagc tgctcgtcaa gctcaaccga gaggacctcc     4260

ttcgaaagca gcgaaccttc gataacggct ccatccccca ccagatccac ctgggtgagc     4320

tccacgccat tctccgaaga caagaggact tctacccctt cctaaaggat aaccgagaga     4380

agatcgagaa gattctcacc ttccgaatcc cctactacgt cggtcccctc gctcgaggta     4440

actcccgatt tgcttggatg acccgaaagt ccgaggagac tatcaccccc tggaactttg     4500

aagaggtagt cgacaagggt gcctccgccc agtctttcat tgagcggatg accaacttcg     4560

ataagaacct ccccaacgag aaggtccttc ccaagcactc tctcctctac gagtacttca     4620

ccgtctacaa cgagctgacc aaggtcaagt acgttaccga gggcatgcga aagcccgctt     4680

tcctctctgg tgagcagaag aaggccattg tcgacctcct gttcaagact aaccgaaaag     4740

tcaccgtcaa gcagctcaag gaagactact tcaagaagat tgagtgcttc gactccgtcg     4800

agatttccgg tgtcgaggac cgattcaacg cctccctcgg cacctaccac gatcttctga     4860

agatcatcaa ggacaaggac tttcttgata acgaggagaa cgaggacatt ctcgaggaca     4920

tcgtcctcac cctcaccctt ttcgaggatc gagagatgat cgaggagcga ctcaagacct     4980

acgcccatct cttcgacgac aaggtcatga agcaactcaa gcgacgacga tacactggct     5040

ggggccgact ttcccgaaag ctcatcaacg gcatccgaga caagcagtct ggcaagacca     5100

tcctggactt cctgaagtcc gacggtttcg ccaaccgaaa cttcatgcag ctcatccacg     5160

acgactctct taccttcaaa gaggatatcc agaaggccca ggtttctggc cagggcgact     5220

ccctccacga gcacattgcc aacctcgccg gatcccccgc catcaaaaag ggtatcctcc     5280

agaccgtcaa ggttgtcgac gaactcgtga aggtcatggg ccgacacaag cccgagaaca     5340

tcgttatcga gatggcccga gagaaccaga ccacccagaa gggtcagaag aactcccgag     5400

agcgaatgaa gcgaatcgaa gagggtatca aggagctcgg ttcccagatt ctcaaggagc     5460

accccgtcga gaacacccag ctccagaacg agaaactcta cctgtactac ctccagaatg     5520

gccgagacat gtacgttgac caggagctcg acatcaaccg actctccgac tacgacgtcg     5580

accacattgt tcctcagtcc ttcctcaagg acgactccat cgacaacaag gttctgaccc     5640

gatctgacaa gaaccgaggt aagtccgaca acgttccctc cgaagaggtc gttaagaaga     5700

tgaagaacta ctggcgacag cttctcaacg ccaaactgat cacccagcga aagtttgaca     5760

acctcaccaa ggccgagcga ggtggtctgt ccgagctgga caaggccggc ttcattaagc     5820

gacagctggt cgagactcga cagatcacca agcacgtcgc ccagatcctc gactcccgaa     5880

tgaacaccaa gtacgacgag aacgacaagc tcatccggga ggtcaaggtc atcaccctga     5940

agtctaagct tgtctccgac ttccgaaagg acttccagtt ctacaaggtc cgagagatca     6000

acaactacca ccacgcccac gacgcctacc tcaacgccgt tgttggtacc gccctcatca     6060

agaagtatcc caagctcgag tccgagttcg tttacggcga ctacaaggtt tacgatgtcc     6120

gaaagatgat tgccaagtcc gagcaggaga tcggtaaggc caccgccaag tactttttct     6180

actccaacat catgaatttc ttcaagaccg agatcactct cgccaacggt gagattcgaa     6240

agcgacccct gattgagact aatggtgaga ctggtgagat cgtctgggat aagggccgag     6300

acttcgccac cgtccgaaag gtcctgtcca tgccccaggt caacattgtc aagaagaccg     6360

aggtccagac cggtggcttc tccaaggagt ccattctccc caagcgaaac tccgacaaac     6420

tcatcgcccg taagaaggac tgggatccga agaagtacgg tggtttcgat tctcccaccg     6480

ttgcctactc cgtcctcgtt gttgctaaag tcgagaaggg taagtctaag aaactcaagt     6540

ccgtgaagga gctactcggt atcaccatca tggagcgatc ttcttttgag aagaacccca     6600

ttgacttcct cgaggccaag ggttacaaag aggtcaagaa ggacctgatt atcaagctgc     6660

ccaagtactc cctctttgag ctcgagaacg gccgaaagcg aatgctggct tccgctggtg     6720

agctgcagaa gggcaacgag ctcgctctgc cctccaagta cgtcaacttc ctctacctgg     6780

cctcccacta cgagaagctc aagggctccc ccgaggacaa cgagcagaag cagctgttcg     6840

ttgagcagca caagcactac ctcgacgaga tcatcgagca gatctccgag ttctccaagc     6900

gagtcatcct cgctgacgcc aaccttgata aggttctctc tgcttacaac aagcaccggg     6960

acaagcccat ccgagagcag gccgagaata tcatccacct cttcactctc accaacctcg     7020

gcgctcctgc tgccttcaag tacttcgaca ccaccattga ccgaaagagg tacacctcca     7080

ccaaggaagt cctcgacgcc accctgatcc accagtccat caccggcctc tacgaaaccc     7140

gaatcgacct ctcccagctc ggcggtgact ctcgagccga ccccaagaag aagcgaaaag     7200

tctaaatatc cgaagatcaa gagcgaagca agttgtaagt ccaggacatg tttcccgccc     7260

acgcgagtga tttataacac ctctcttttt tgacacccgc tcgccttgaa attcatgtca     7320

cataaattat agtcaacgac gtttgaataa cttgtcttgt agttcgatga tgatcatatg     7380

attacattaa tagtaattac tgtatttgat atatatacta attacaatag tacatattag     7440

aacatacaat agttagtgcc gtgaagtggc ttaaaatacc gcgagtcgat tacgtaatat     7500

tattacctct tgcccatcga acgtacaagt actcctctgt tctctccttc ctttgctttg     7560

tgcacgaaga actgcggtca ggtgacacaa ctttttccat ctcagggtgt gtcgcgtgtg     7620

cttcatccaa actttagttg gggttcgggt tcgcgcgaga tgatcacgtg ccctgatttg     7680

gtgtcgtccc ccgtcgcgct gcgcacgtga tttatttatt tccggtggct gctgtctacg     7740

cggggccttc tctgcccttc tgtttcaacc ttcgggcggt tctcgtaacc agcagtagca     7800

atccatttcg aaactcaaag agctaaaaac gttaaacctc agcagtcgct cgacgaatgg     7860

gctgcggttg ggaagcccac gaggcctata gccagagcct cgagttgaca ggagcccaga     7920

cgccttttcc aacggcaact tttatataaa atggcaatgt attcatgcaa ttgcggccgt     7980

gtcaggttgg agacactgga ccacactctc cattgcttcc tgaggagatg gatcattgct     8040

agtgcatcta cgcgcagcaa tcccgcaagc tcgacaaccg tagatgggct ttggtgggcc     8100

aatcaattac gcaacccgca cgttaaattg tatgaggaag gaaggccacg gtacaaagtg     8160

ggtggtcttc acccagtggt tgttggtggc gtcatgcaga ccatgcattg gggatagcac     8220

agggttgggg tgtcttgtgg actcaatggg tgaaaggaga tggaaaaggg cggtgaaaag     8280

tggtagaatc gaaatccctg acgtcaattt ataaagtaaa atgcgtttct gccattttgc     8340

tcccctcctt ctttcgcaat cgcctcccca aaagttgtcg tggcagtaca catgcttgca     8400

tacaatgaag ctaatccggc ttgctcagta gttgctatat ccaggcatgg tgtgaaaccc     8460

ctcaaagtat atataggagc ggtgagcccc agtctggggt cttttctctc catctcaaaa     8520

ctactttctc acaatgaggc cactgatgag tccgtgagga cgaaacgagt aagctcgtct     8580

ggcctgttga gtcaacccgg ttttagagct agaaatagca agttaaaata aggctagtcc     8640

gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttg gccggcatgg tcccagcctc     8700

ctcgctggcg ccggctgggc aacatgcttc ggcatggcga atgggactaa acttcgagct     8760

aatccagtag cttacgttac ccaggggcag gtcaactggc tagccacgag tctgtcccag     8820

gtcgcaattt agtgtaataa acaatatata tattgagtct aaagggaatt gtagctattg     8880

tgattgtgtg attttcgtct tgctggttct tattgtgtcc cattcgtttc atcctgatga     8940

ggacccctgg aaccggtgtt ttcttagtct ctgcaatcgc tagtcttgtt gctatgacag     9000

ttgcgtcgac actattcagg tcatctatcg gttattctga tattataata cctccggatc     9060

gatgtacctg atttatactt gcagcaatgt ttacttctta tcgcgataca cgaatgtgat     9120

acggatcaaa gtaagcagga ctacgataag ataacgaatg cggtgcagtc catgtcgatt     9180

aggtatagat acatttattt tgtgttatgt tacattttgg ggggatactg tcctacttgt     9240

agtacctact tgtagtggcg cgtctattcc tttgccctcg gacgagtgct ggggcgtcgg     9300

tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg     9360

gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc     9420

tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga     9480

ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc     9540

tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg     9600

gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg     9660

cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact     9720

tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg     9780

acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg     9840

aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc     9900

gtctggctaa gatcggccgc agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca     9960

gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg    10020

caataggtca ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg    10080

gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt    10140

acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc    10200

tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca    10260

gacgtcgcgg tgagttcagg ctttttcata tgggtacctg agaacatttt tgtgtctagg    10320

tgtttgtgtt tggactgcga tcagtgaaga aaagaagagg aaaaattgtg caagaaattt    10380

tgctttcaag acttggctga tgcagcaggg taactctggg acacagacct atgtttgtgg    10440

ttaaactcaa tgcacgtggt acgtgcgtgg agcgcttacc catccaaggg tgtggacatg    10500

gaaccgacgg tccgtggagt tgtgtaatgt cattttggcg actcttgaag caaggctata    10560

aaaaaattgt gtggcttgag tcttatcgag ctcggtcact acaagagtta atcttcctgt    10620

ctcaggcaga caggtcaggc agggttactt ttgggtgtgc tgtaactcac tgtatggccg    10680

ttagtgcgca tagacgttgt acatactgga ccgaattgta gcgtgctcaa tagggccaat    10740

aaagctattg tagggatccg aattttcaga acctaattta tctgttaccc ggcctgtggc    10800

tcgcacagct taaaaatggt caaactttcc ccttcttgtc tttttttcct cacattcatc    10860

aggttcttgt cttgatcttt caagtgagta ttaattaccg accttggttc ttcattggga    10920

gagcattgga agccgtggtg cagcaaccac aaaacggttc ttccccttcg ataccttctt    10980

gcctgccttt caatacaagt cggctcgatt agcggtggtc gcccccgcca gcggagaaca    11040

tggaactaac ccagaatgag agctaagtgg agaaagaaga gagtcagacg actcaagcga    11100

aagcgccgca aggtccgagc tcgatccaaa taagcggttt ttaacggaga tttaacacta    11160

aatcgaagaa cttttcccgt ttcatttgcg aatgagctcg ttaacaaaat cccccagttt    11220

ttttatccag ctgtaaggat tgacattagt aatgaattat tgtttggtat atttaaatct    11280

gtagttcctt tctgtccgtg tcggcaactg tcgtactcgt gatttacttg tattgacgaa    11340

tacttactgt agcgcactct gctgctactg gtcgtaagga tgtgctattt cggtgtatgg    11400

tgggtttttt gggggtcgga accgaagact gttacacggg cacggctcgt tgtgtacacg    11460

cacagagctc ttgcgagtca tgttgtagct agctcgtcgt gttcaggaac tgttcgatgg    11520

ttcggagaga gtcgccgccc agaacatacg cgcaccgatg tcagcagaca gccttattac    11580

aagtatattc aagcaagtat atccgtaggg tgcgggtgat ttggatctaa ggttcgtact    11640

caacactcac gagcagcttg cctatgttac atccttttat cagacataac ataattggag    11700

tttacttaca cacggggtgt acctgtatga gcaccaccta caattgtagc actggtactt    11760

gtacaaagaa tttattcgta cgaatcacag ggacggccgc cctcaccgaa ccagcgaata    11820

cctcagcggt cccctgcagt gactcaacaa agcgatatga acatcttgcg atggtatcct    11880

gctgatagtt tttactgtac aaacacctgt gtagctcctt ctagcatttt taagttattc    11940

acacctcaag gggagggata aattaaataa attccaaaag cgaagatcga gaaactaaat    12000

taaaattcca aaaacgaagt tggaacacaa ccccccgaaa aaaaacaaca aacaaaaaac    12060

ccaacaaaat aaacaaaaac aaaataaata tataactacc agtatctgac taaaagttca    12120

aatactcgta cttacaacaa atagaaatga gccggccaaa attctgcaga aaaaaatttc    12180

aaacaagtac tggtataatt aaattaaaaa acacatcaaa gtatcataac gttagttatt    12240

ttattttatt taataaaaga aaacaacaag atgggctcaa aactttcaac ttatacgata    12300

cataccaaat aacaatttag tatttatcta agtgcttttc gtagataatg gaatacaaat    12360

ggatatccag agtatacaca tggatagtat acactgacac gacaattctg tatctcttta    12420

tgttaactac tgtgaggcat taaatagagc ttgatatata aaatgttaca tttcacagtc    12480

tgaacttttg cagattacct aatttggtaa gatattaatt atgaactgaa agttgatggc    12540

atccctaaat ttgatgaaag atgaaattgt aaatgaggtg gtaaaagagc tacagtcgtt    12600

ttgttttgag ataccatcat ctctaacgaa atatctatta aaaatctcag tgtgatcatg    12660

agtcattgcc atcctggaaa atgtcatcat ggctgatatt tctaactgtt tacttgagat    12720

aaatatatat ttacaagaac ttcccttgaa attaatttag atataaaatg tttgcgggca    12780

agttactacg aggaataaat tatatctaga                                     12810


<210>  158
<211>  2142
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the GFP expression cassette (Yl_HSP.pro - 
       A.vic_eGFP ORF - Yl_GPD.ter) flanked by 50-bp genomic DNA 
       sequences on either side for targeted integration in the INT05 
       locus

<400>  158
tgtgaccttg taaacaacaa cccgacttta cagtaacgct tggacacact gtgcaatcac       60

atgttgctac tgtacctgct gtggaccacg cacggcggaa cgtaccgtac aaatattttc      120

ttgctcacat gactctctct cggccgcgca cgccggtggc aaattgctct tgcattggct      180

ctgtctctag acgtccaaac cgtccaaagt ggcagggtga cgtgatgcga cgcacgaagg      240

agatggcccg gtggcgagga accggacacg gcgagccggc gggaaaaaag gcggaaaacg      300

aaaagcgaag ggcacaatct gacggtgcgg ctgccaccaa cccaaggagg ctattttggg      360

tcgctttcca tttcacattc gccctcaatg gccactttgc ggtggtgaac atggtttctg      420

aaacaacccc ccagaattag agtatattga tgtgtttaag attgggttgc tatttggcca      480

ttgtggggga gggtagcgac gtggaggaca ttccagggcg aattgagcct agaaagtggt      540

agcattccaa ccgtctaagt cgtccgaatt gatcgctata actatcacct ctctcacatg      600

tctacttccc caaccaacat ccccaacctc ccccacacta aagttcacgc caataatgta      660

ggcactcttt ctgggtgtgg gacagcagag caatacggag gggagattac acaacgagcc      720

acaattgggg agatggtagc catctcactc gacccgtcga cttttggcaa cgctcaatta      780

cccaccaaat ttgggctgga gttgagggga ccgtgttcca gcgctgtagg accagcaaca      840

cacacggtat caacagcaac caacgccccc gctaatgcac ccagtactgc gcaggtgtgg      900

gccaggtgcg ttccagatgc gagttggcga accctaagcc gacagtgtac tttttgggac      960

gggcagtagc aatcgtgggc ggaaaccccg gtgtatataa aggggtggag aggacggatt     1020

attagcacca acacacacac ttatactaca atggagggtg atattcacgg tgcttctaag     1080

ggtgaggagc ttttcaccgg cgttgtccct attctcgttg agcttgatgg tgatgtcaac     1140

ggtcacaagt tttctgtctc tggtgagggt gagggtgatg ctacttacgg taagctcacc     1200

ctcaagttta tttgtaccac cggcaagctc cccgttccct ggcctaccct cgttaccact     1260

ctcacctacg gtgttcagtg tttttcccga taccccgatc acatgaagcg acacgacttt     1320

ttcaagtctg ccatgcccga gggttacgtt caggagcgaa ctatttcttt caaggatgac     1380

ggtaactaca agacccgagc tgaggttaag tttgagggtg ataccctcgt taaccgaatc     1440

gagcttaagg gtattgattt caaggaggat ggtaacattc tcggtcacaa gctggagtac     1500

aactacaact ctcacaacgt ttacatcacc gctgacaagc agaagaacgg tatcaaggct     1560

aacttcaaga ttcgacacaa cattgaggat ggttccgttc agcttgctga ccactaccag     1620

cagaacactc ccattggcga tggccctgtc ctcctccccg acaaccacta cctctctacc     1680

cagtctgccc tttctaagga ccccaacgag aagcgagatc acatggtcct ccttgagttc     1740

gttaccgctg ctggtattac tcacggtatg gatgagctct acaagtaaat aaatatccga     1800

agatcaagag cgaagcaagt tgtaagtcca ggacatgttt cccgcccacg cgagtgattt     1860

ataacacctc tcttttttga cacccgctcg ccttgaaatt catgtcacat aaattatagt     1920

caacgacgtt tgaataactt gtcttgtagt tcgatgatga tcatatgatt acattaatag     1980

taattactgt atttgatata tatactaatt acaatagtac atattagaac atacaatagt     2040

tagtgccgtg aagtggctta aaataccgcg agtcgattac gtaatattat tacatcaata     2100

tatcggacca atatatcgga ccacggtcgc cgccgaatcg cc                        2142


<210>  159
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to confirm integration 
       of the GFP expression cassette in the INT05 locus in the Yarrowia
       genome

<400>  159
cattgatctt actgtcgaat gtac                                              24


<210>  160
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to confirm integration 
       of the GFP expression cassette in the INT05 locus in the Yarrowia
       genome

<400>  160
ttggcgtagg tgccgatgta g                                                 21


<210>  161
<211>  5844
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of plasmid pSTV077

<400>  161
agatcttaat taagcttggc gcgcctccgg atcgatgtac ctgatttata cttgcagcaa       60

tgtttacttc ttatcgcgat acacgaatgt gatacggatc aaagtaagca ggactacgat      120

aagataacga atgcggtgca gtccatgtcg attaggtata gatacattta ttttgtgtta      180

tgttacattt tggggggata ctgtcctact tgtagtacct acttgtagtg gcgcgtctat      240

tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag tacttctaca      300

cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg      360

gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg      420

ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata cgcccggagc      480

cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg ctgctccata      540

caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga atccccgaac      600

atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag gacattgttg      660

gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc ccaaagcatc      720

agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac agtttgccag      780

tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt gtattgaccg      840

attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc cgcagcgatc      900

gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc aggcaggtct      960

tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc gctgaattcc     1020

ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg ataaacataa     1080

cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc acgccctcct     1140

acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag gtcggagacg     1200

ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc aggctttttc     1260

atatgggtac ctgagaacat ttttgtgtct aggtgtttgt gtttggactg cgatcagtga     1320

agaaaagaag aggaaaaatt gtgcaagaaa ttttgctttc aagacttggc tgatgcagca     1380

gggtaactct gggacacaga cctatgtttg tggttaaact caatgcacgt ggtacgtgcg     1440

tggagcgctt acccatccaa gggtgtggac atggaaccga cggtccgtgg agttgtgtaa     1500

tgtcattttg gcgactcttg aagcaaggct ataaaaaaat tgtgtggctt gagtcttatc     1560

gagctcggtc actacaagag ttaatcttcc tgtctcaggc agacaggtca ggcagggtta     1620

cttttgggtg tgctgtaact cactgtatgg ccgttagtgc gcatagacgt tgtacatact     1680

ggaccgaatt gtagcgtgct caatagggcc aataaagcta ttgtagggat ccgaattttc     1740

agaacctaat ttatctgtta cccggcctgt ggctcgcaca gcttaaaaat ggtcaaactt     1800

tccccttctt gtcttttttt cctcacattc atcaggttct tgtcttgatc tttcaagtga     1860

gtattaatta ccgaccttgg ttcttcattg ggagagcatt ggaagccgtg gtgcagcaac     1920

cacaaaacgg ttcttcccct tcgatacctt cttgcctgcc tttcaataca agtcggctcg     1980

attagcggtg gtcgcccccg ccagcggaga acatggaact aacccagaat gagagctaag     2040

tggagaaaga agagagtcag acgactcaag cgaaagcgcc gcaaggtccg agctcgatcc     2100

aaataagcgg tttttaacgg agatttaaca ctaaatcgaa gaacttttcc cgtttcattt     2160

gcgaatgagc tcgttaacaa aatcccccag tttttttatc cagctgtaag gattgacatt     2220

agtaatgaat tattgtttgg tatatttaaa tctgtagttc ctttctgtcc gtgtcggcaa     2280

ctgtcgtact cgtgatttac ttgtattgac gaatacttac tgtagcgcac tctgctgcta     2340

ctggtcgtaa ggatgtgcta tttcggtgta tggtgggttt tttgggggtc ggaaccgaag     2400

actgttacac gggcacggct cgttgtgtac acgcacagag ctcttgcgag tcatgttgta     2460

gctagctcgt cgtgttcagg aactgttcga tggttcggag agagtcgccg cccagaacat     2520

acgcgcaccg atgtcagcag acagccttat tacaagtata ttcaagcaag tatatccgta     2580

gggtgcgggt gatttggatc taaggttcgt actcaacact cacgagcagc ttgcctatgt     2640

tacatccttt tatcagacat aacataattg gagtttactt acacacgggg tgtacctgta     2700

tgagcaccac ctacaattgt agcactggta cttgtacaaa gaatttattc gtacgaatca     2760

cagggacggc cgccctcacc gaaccagcga atacctcagc ggtcccctgc agtgactcaa     2820

caaagcgata tgaacatctt gcgatggtat cctgctgata gtttttactg tacaaacacc     2880

tgtgtagctc cttctagcat ttttaagtta ttcacacctc aaggggaggg ataaattaaa     2940

taaattccaa aagcgaagat cgagaaacta aattaaaatt ccaaaaacga agttggaaca     3000

caaccccccg aaaaaaaaca acaaacaaaa aacccaacaa aataaacaaa aacaaaataa     3060

atatataact accagtatct gactaaaagt tcaaatactc gtacttacaa caaatagaaa     3120

tgagccggcc aaaattctgc agaaaaaaat ttcaaacaag tactggtata attaaattaa     3180

aaaacacatc aaagtatcat aacgttagtt attttatttt atttaataaa agaaaacaac     3240

aagatgggct caaaactttc aacttatacg atacatacca aataacaatt tagtatttat     3300

ctaagtgctt ttcgtagata atggaataca aatggatatc cagagtatac acatggatag     3360

tatacactga cacgacaatt ctgtatctct ttatgttaac tactgtgagg cattaaatag     3420

agcttgatat ataaaatgtt acatttcaca gtctgaactt ttgcagatta cctaatttgg     3480

taagatatta attatgaact gaaagttgat ggcatcccta aatttgatga aagatgaaat     3540

tgtaaatgag gtggtaaaag agctacagtc gttttgtttt gagataccat catctctaac     3600

gaaatatcta ttaaaaatct cagtgtgatc atgagtcatt gccatcctgg aaaatgtcat     3660

catggctgat atttctaact gtttacttga gataaatata tatttacaag aacttccctt     3720

gaaattaatt tagatataaa atgtttgcgg gcaagttact acgaggaata aattatatct     3780

agaggttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg     3840

gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga     3900

aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg     3960

gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag     4020

aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc     4080

gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg     4140

ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt     4200

cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc     4260

ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc     4320

actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg     4380

tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca     4440

gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc     4500

ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat     4560

cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt     4620

ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt     4680

tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc     4740

agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc     4800

gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata     4860

ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg     4920

gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc     4980

cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct     5040

gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa     5100

cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt     5160

cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca     5220

ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac     5280

tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca     5340

acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt     5400

tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc     5460

actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca     5520

aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata     5580

ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc     5640

ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc     5700

cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat     5760

aggcgtatca cgaggccctt tcgtctggcc taggaagcga cttccaatcg ctttgcatat     5820

ccagtaccac acccacaggc gttt                                            5844


<210>  162
<211>  1000
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Yarrowia Yl_HSP promoter

<400>  162
gtgcaatcac atgttgctac tgtacctgct gtggaccacg cacggcggaa cgtaccgtac       60

aaatattttc ttgctcacat gactctctct cggccgcgca cgccggtggc aaattgctct      120

tgcattggct ctgtctctag acgtccaaac cgtccaaagt ggcagggtga cgtgatgcga      180

cgcacgaagg agatggcccg gtggcgagga accggacacg gcgagccggc gggaaaaaag      240

gcggaaaacg aaaagcgaag ggcacaatct gacggtgcgg ctgccaccaa cccaaggagg      300

ctattttggg tcgctttcca tttcacattc gccctcaatg gccactttgc ggtggtgaac      360

atggtttctg aaacaacccc ccagaattag agtatattga tgtgtttaag attgggttgc      420

tatttggcca ttgtggggga gggtagcgac gtggaggaca ttccagggcg aattgagcct      480

agaaagtggt agcattccaa ccgtctaagt cgtccgaatt gatcgctata actatcacct      540

ctctcacatg tctacttccc caaccaacat ccccaacctc ccccacacta aagttcacgc      600

caataatgta ggcactcttt ctgggtgtgg gacagcagag caatacggag gggagattac      660

acaacgagcc acaattgggg agatggtagc catctcactc gacccgtcga cttttggcaa      720

cgctcaatta cccaccaaat ttgggctgga gttgagggga ccgtgttcca gcgctgtagg      780

accagcaaca cacacggtat caacagcaac caacgccccc gctaatgcac ccagtactgc      840

gcaggtgtgg gccaggtgcg ttccagatgc gagttggcga accctaagcc gacagtgtac      900

tttttgggac gggcagtagc aatcgtgggc ggaaaccccg gtgtatataa aggggtggag      960

aggacggatt attagcacca acacacacac ttatactaca                           1000


<210>  163
<211>  742
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Aequorea victoria eGFP gene (A. vic_eGFP 
       ORF)

<400>  163
atggagggtg atattcacgg tgcttctaag ggtgaggagc ttttcaccgg cgttgtccct       60

attctcgttg agcttgatgg tgatgtcaac ggtcacaagt tttctgtctc tggtgagggt      120

gagggtgatg ctacttacgg taagctcacc ctcaagttta tttgtaccac cggcaagctc      180

cccgttccct ggcctaccct cgttaccact ctcacctacg gtgttcagtg tttttcccga      240

taccccgatc acatgaagcg acacgacttt ttcaagtctg ccatgcccga gggttacgtt      300

caggagcgaa ctatttcttt caaggatgac ggtaactaca agacccgagc tgaggttaag      360

tttgagggtg ataccctcgt taaccgaatc gagcttaagg gtattgattt caaggaggat      420

ggtaacattc tcggtcacaa gctggagtac aactacaact ctcacaacgt ttacatcacc      480

gctgacaagc agaagaacgg tatcaaggct aacttcaaga ttcgacacaa cattgaggat      540

ggttccgttc agcttgctga ccactaccag cagaacactc ccattggcga tggccctgtc      600

ctcctccccg acaaccacta cctctctacc cagtctgccc tttctaagga ccccaacgag      660

aagcgagatc acatggtcct ccttgagttc gttaccgctg ctggtattac tcacggtatg      720

gatgagctct acaagtaaat aa                                               742


<210>  164
<211>  300
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of Yarrowia Yl_GPD terminator

<400>  164
atatccgaag atcaagagcg aagcaagttg taagtccagg acatgtttcc cgcccacgcg       60

agtgatttat aacacctctc ttttttgaca cccgctcgcc ttgaaattca tgtcacataa      120

attatagtca acgacgtttg aataacttgt cttgtagttc gatgatgatc atatgattac      180

attaatagta attactgtat ttgatatata tactaattac aatagtacat attagaacat      240

acaatagtta gtgccgtgaa gtggcttaaa ataccgcgag tcgattacgt aatattatta      300


<210>  165
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the forward primer to amplify the edited 
       GFP ORF from the Yarrowia genome

<400>  165
atggagggtg atattcacgg tg                                                22


<210>  166
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the reverse primer to amplify the edited 
       GFP ORF

from the Yarrowia genome

<400>  166
ttacttgtag agctcatcca tac                                               23


<210>  167
<211>  6
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of 6 bp inverted repeat of the KU70 genomic 
       target

<400>  167
cgatat                                                                   6


<210>  168
<211>  6
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of 6 bp inverted repeat of the INT05 genomic 
       target

<400>  168
aggcca                                                                   6


<210>  169
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of the 20-bp genomic target sequence of the 
       INT05 locus

<400>  169
tggcctgttg agtcaacccg                                                   20


<210>  170
<211>  1587
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC DNA fragment 1, comprising a guide 
       RNA expression cassette (sgRNA) for targeting Cas9 to the GFP 
       gene and donor DNA of 100-bp, which encodes for the full knock 
       out of the GFP ORF, on the 3' side

<400>  170
acgaagaact gcggtcaggt gacacaactt tttccatctc agggtgtgtc gcgtgtgctt       60

catccaaact ttagttgggg ttcgggttcg cgcgagatga tcacgtgccc tgatttggtg      120

tcgtcccccg tcgcgctgcg cacgtgattt atttatttcc ggtggctgct gtctacgcgg      180

ggccttctct gcccttctgt ttcaaccttc gggcggttct cgtaaccagc agtagcaatc      240

catttcgaaa ctcaaagagc taaaaacgtt aaacctcagc agtcgctcga cgaatgggct      300

gcggttggga agcccacgag gcctatagcc agagcctcga gttgacagga gcccagacgc      360

cttttccaac ggcaactttt atataaaatg gcaatgtatt catgcaattg cggccgtgtc      420

aggttggaga cactggacca cactctccat tgcttcctga ggagatggat cattgctagt      480

gcatctacgc gcagcaatcc cgcaagctcg acaaccgtag atgggctttg gtgggccaat      540

caattacgca acccgcacgt taaattgtat gaggaaggaa ggccacggta caaagtgggt      600

ggtcttcacc cagtggttgt tggtggcgtc atgcagacca tgcattgggg atagcacagg      660

gttggggtgt cttgtggact caatgggtga aaggagatgg aaaagggcgg tgaaaagtgg      720

tagaatcgaa atccctgacg tcaatttata aagtaaaatg cgtttctgcc attttgctcc      780

cctccttctt tcgcaatcgc ctccccaaaa gttgtcgtgg cagtacacat gcttgcatac      840

aatgaagcta atccggcttg ctcagtagtt gctatatcca ggcatggtgt gaaacccctc      900

aaagtatata taggagcggt gagccccagt ctggggtctt ttctctccat ctcaaaacta      960

ctttctcaca atggtattgc tgatgagtcc gtgaggacga aacgagtaag ctcgtccaat     1020

acccttaagc tcgattgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1080

atcaacttga aaaagtggca ccgagtcggt gcttttggcc ggcatggtcc cagcctcctc     1140

gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggactaaact tcgagctaat     1200

ccagtagctt acgttaccca ggggcaggtc aactggctag ccacgagtct gtcccaggtc     1260

gcaatttagt gtaataaaca atatatatat tgagtctaaa gggaattgta gctattgtga     1320

ttgtgtgatt ttcgtcttgc tggttcttat tgtgtcccat tcgtttcatc ctgatgagga     1380

cccctggaac cggtgttttc ttagtctctg caatcgctag tcttgttgct atgacagttg     1440

cgtcgacact attcaggtca tctatcggtt attctgatat tataataggg aaacatgtcc     1500

tggacttaca acttgcttcg ctcttgatct tcggatagta gtataagtgt gtgtgttggt     1560

gctaataatc cgtcctctcc acccctt                                         1587


<210>  171
<211>  1587
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of CTEC DNA fragment 2, comprising a guide 
       RNA expression cassette (sgRNA) for targeting Cas9 to the GFP 
       gene and donor DNA of 100-bp, which encodes a base deletion in 
       the PAM sequence, changing it from CGG to CG, on the 3' side

<400>  171
acgaagaact gcggtcaggt gacacaactt tttccatctc agggtgtgtc gcgtgtgctt       60

catccaaact ttagttgggg ttcgggttcg cgcgagatga tcacgtgccc tgatttggtg      120

tcgtcccccg tcgcgctgcg cacgtgattt atttatttcc ggtggctgct gtctacgcgg      180

ggccttctct gcccttctgt ttcaaccttc gggcggttct cgtaaccagc agtagcaatc      240

catttcgaaa ctcaaagagc taaaaacgtt aaacctcagc agtcgctcga cgaatgggct      300

gcggttggga agcccacgag gcctatagcc agagcctcga gttgacagga gcccagacgc      360

cttttccaac ggcaactttt atataaaatg gcaatgtatt catgcaattg cggccgtgtc      420

aggttggaga cactggacca cactctccat tgcttcctga ggagatggat cattgctagt      480

gcatctacgc gcagcaatcc cgcaagctcg acaaccgtag atgggctttg gtgggccaat      540

caattacgca acccgcacgt taaattgtat gaggaaggaa ggccacggta caaagtgggt      600

ggtcttcacc cagtggttgt tggtggcgtc atgcagacca tgcattgggg atagcacagg      660

gttggggtgt cttgtggact caatgggtga aaggagatgg aaaagggcgg tgaaaagtgg      720

tagaatcgaa atccctgacg tcaatttata aagtaaaatg cgtttctgcc attttgctcc      780

cctccttctt tcgcaatcgc ctccccaaaa gttgtcgtgg cagtacacat gcttgcatac      840

aatgaagcta atccggcttg ctcagtagtt gctatatcca ggcatggtgt gaaacccctc      900

aaagtatata taggagcggt gagccccagt ctggggtctt ttctctccat ctcaaaacta      960

ctttctcaca atggtattgc tgatgagtcc gtgaggacga aacgagtaag ctcgtccaat     1020

acccttaagc tcgattgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     1080

atcaacttga aaaagtggca ccgagtcggt gcttttggcc ggcatggtcc cagcctcctc     1140

gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggactaaact tcgagctaat     1200

ccagtagctt acgttaccca ggggcaggtc aactggctag ccacgagtct gtcccaggtc     1260

gcaatttagt gtaataaaca atatatatat tgagtctaaa gggaattgta gctattgtga     1320

ttgtgtgatt ttcgtcttgc tggttcttat tgtgtcccat tcgtttcatc ctgatgagga     1380

cccctggaac cggtgttttc ttagtctctg caatcgctag tcttgttgct atgacagttg     1440

cgtcgacact attcaggtca tctatcggtt attctgatat tataatatcc agcttgtgac     1500

cgagaatgtt accatcctcc ttgaaatcaa tacccttaag ctcgattcgt taacgagggt     1560

atcaccctca aacttaacct cagctcg                                         1587


