LOCUS Exported 10793 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses nCas9(H840A) in yeast cells. ACCESSION . VERSION . KEYWORDS pRS415_pGal-nCas9(H840A) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10793) AUTHORS Nishida K, Arazoe T, Yachie N, Banno S, Kakimoto M, Tabata M, Mochizuki M, Miyabe A, Araki M, Hara KY, Shimatani Z, Kondo A TITLE Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. JOURNAL Science. 2016 Aug 4. pii: aaf8729. PUBMED 27492474 REFERENCE 2 (bases 1 to 10793) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10793 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 70..573 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 610..714 /gene="bla" /label=AmpR promoter CDS 715..1575 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(933..952) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1746..2334 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2235..2254 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2488..2505 /label=L4440 /note="L4440 vector, forward primer" promoter 2658..2688 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2696..2712 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2701..2723 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2720..2736 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2720..2736 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2755..2775 /label=T3 /note="T3 promoter, forward primer" promoter 2757..2775 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind 3184..3207 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" primer_bind 3228..3244 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 3228..3244 /label=pBluescriptSK /note="For pBluescript vector" CDS 3258..7361 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(H840A) /note="generates RNA-guided single strand nicks in DNA due to the H840A mutation in the HNH catalytic domain" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 7374..7394 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 7410..7657 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(7426..7444) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(7675..7694) /label=T7 /note="T7 promoter, forward primer" promoter complement(7676..7694) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7704..7721) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7704..7720) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7713..7735) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7865..8320 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7952..7971) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8162..8183 /label=F1ori-F /note="F1 origin, forward primer" promoter 8620..9024 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 9037..10131 /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" primer_bind complement(10624..10643) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 10743..10765 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata 121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt 181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa 241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata 301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct 361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata 421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact 481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa 541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg 601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact 2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg 2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt 2761 aaccctcact aaagggaaca aaagctggag ctcgtgacag ccctccgaag gaagactctc 2821 ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc 2881 actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat 2941 tggcagtaac ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat 3001 gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt 3061 tttgatctat taacggatat ataaatgcaa aaactgcata accactttaa ctaatacttt 3121 caacattttc ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt 3181 gttaatatac ctctatactt taacgtcaag gagaaaaaac cccggattct agaactagtg 3241 gatcccccgg gaaaaaaatg gacaagaagt actccattgg gctcgatatc ggcacaaaca 3301 gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc 3361 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact 3421 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca 3481 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg 3541 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc 3601 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca 3661 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct 3721 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga 3781 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc 3841 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta 3901 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga 3961 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta 4021 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc 4081 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga 4141 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag 4201 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc 4261 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt 4321 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat 4381 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca 4441 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc 4501 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag 4561 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc 4621 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc 4681 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga 4741 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt 4801 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga 4861 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga 4921 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt 4981 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc 5041 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca 5101 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac 5161 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc 5221 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga 5281 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc 5341 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg 5401 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa 5461 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata 5521 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga 5581 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa 5641 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact 5701 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg 5761 actacgacgt ggatgctatc gtgccccagt cttttctcaa agatgattct attgataata 5821 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag 5881 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac 5941 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg 6001 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc 6061 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag 6121 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg 6181 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca 6241 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag 6301 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta 6361 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg 6421 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg 6481 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg 6541 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga 6601 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg 6661 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta 6721 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg 6781 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca 6841 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg 6901 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt 6961 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga 7021 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg 7081 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca 7141 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc 7201 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc 7261 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc 7321 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga 7381 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc 7441 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta 7501 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat 7561 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct 7621 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat 7681 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac 7741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat 7801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg 7861 cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 7921 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 7981 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 8041 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 8101 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 8161 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 8221 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 8281 tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc 8341 ttacgcatct gtgcggtatt tcacaccgca tatcgacggt cgaggagaac ttctagtata 8401 tccacatacc taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc 8461 cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt 8521 atatttatag gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg 8581 tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta 8641 tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc aacataacga 8701 gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat 8761 ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag 8821 gtgagaggcc ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg 8881 catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg 8941 gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat 9001 atatttcaag gatataccat tctaatgtct gcccctatgt ctgcccctaa gaagatcgtc 9061 gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa 9121 gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt 9181 gctgctatcg atgctacagg tgtcccactt ccagatgagg cgctggaagc ctccaagaag 9241 gttgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga 9301 cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca 9361 tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa 9421 ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag 9481 gaagacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa 9541 agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg 9601 tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa 9661 accatcaaga acgaattccc tacattgaag gttcaacatc aattgattga ttctgccgcc 9721 atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt 9781 ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct 9841 gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac 9901 ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat cttgtctgct 9961 gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca 10021 gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc 10081 accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta aaaagattct 10141 ctttttttat gatatttgta cataaacttt ataaatgaaa ttcataatag aaacgacacg 10201 aaattacaaa atggaatatg ttcatagggt agacgaaact atatacgcaa tctacataca 10261 tttatcaaga aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa ttgatactaa 10321 tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac aaggaggagg 10381 gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc taatttatat 10441 attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg 10501 accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt 10561 ccctcaagaa ttttactctg tcagaaacgg ccttacgacg tagtcgatat ggtgcactct 10621 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 10681 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 10741 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga //