LOCUS Exported 11826 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI sites. LEU2 marker. ACCESSION . VERSION . KEYWORDS bRA90 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11826) AUTHORS Anand R, Beach A, Li K, Haber J TITLE Rad51-mediated double-strand break repair and mismatch correction of divergent substrates. JOURNAL Nature. 2017 Apr 20;544(7650):377-380. doi: 10.1038/nature22046. Epub 2017 Apr 12. PUBMED 28405019 REFERENCE 2 (bases 1 to 11826) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11826 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS complement(783..1877) /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" promoter complement(1878..2285) /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 3323..7426 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 7439..7459 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7463..7483 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7487..7507 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7508..7573 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" terminator 7592..7839 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(7608..7626) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" rep_origin complement(7899..8354) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8036..8057) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8248..8267 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8480..8502 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8494..8511 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8495..8511 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 8518..8537 /label=T7 /note="T7 promoter, forward primer" promoter 8518..8536 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 8569..8585 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 8569..8585 /label=pBluescriptSK /note="For pBluescript vector" promoter 8617..8885 /label=SNR52 promoter /note="promoter for the S. cerevisiae small nucleolar RNA gene SNR52" misc_RNA 8913..8987 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 8992..9011 /label=SUP4 terminator /note="transcription terminator for the S. cerevisiae SUP4 tRNA gene" promoter complement(9053..9071) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(9092..9108) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9092..9108) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9105..9127) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9116..9132 /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)." promoter complement(9140..9170) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9185..9206 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(9323..9340) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9494..10082) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9574..9593) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10253..11113) /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 10876..10895 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(11114..11218) /gene="bla" /label=AmpR promoter misc_feature 11255..11758 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind 11799..11817 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcgaattcg 241 agctcgttta aactggatgg cggcgttagt atcgaatcga cagcagtata gcgaccagca 301 ttcacatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 361 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 421 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatataaa 481 ttaggaatca tagtttcatg attttctgtt acacctaact ttttgtgtgg tgccctcctc 541 cttgtcaata ttaatgttaa agtgcaattc tttttcctta tcacgttgag ccattagtat 601 caatttgctt acctgtattc ctttacatcc tcctttttct ccttcttgat aaatgtatgt 661 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 721 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 781 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 841 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 901 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 961 aagatagtgg cgatagggtt gaccttattc tttggcaaat ctggagcaga accgtggcat 1021 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 1081 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1141 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1201 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1261 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1321 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1381 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1441 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1501 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1561 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1621 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc agccttcttg 1681 gaggcttcca gcgcctcatc tggaagtgga acacctgtag catcgatagc agcaccacca 1741 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1801 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1861 ttcttagggg cagacattag aatggtatat ccttgaaata tatatatata tattgctgaa 1921 atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat tggaaaaaac 1981 aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat ttagtcatga 2041 acgcttctct attctatatg aaaagccggt tccggcgctc tcacctttcc tttttctccc 2101 aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc 2161 cagtcatcga atttgattct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa 2221 aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca 2281 cagttgtatt ctgggcctcc atgtcgctgg ccgggtgacc cggcggggaa agtaccttca 2341 aagaatgggg tcttatcttg ttttgcaagt accactgagc aggataataa tagaaatgat 2401 aatatactat agtagagata acgtcgatga cttcccatac tgtaattgct tttagttgtg 2461 tatttttagt gtgcaagttt ctgtaaatcg attaattttt ttttctttcc tctttttatt 2521 aaccttaatt tttattttag attcctgact tcaactcaag acgcacagat attataacat 2581 ctgcataata ggcatttgca agaattactc gtgagtaagg aaagagtgag gaactatcgc 2641 atacctgcat ttaaagatgc cgatttgggc gcgaatcctt tattttggct tcaccctcat 2701 actattatca gggccagaaa aaggaagtgt ttccctcctt cttgaattga tgttaccctc 2761 ataaagcacg tggcctctta tcgagaaaga aattaccgtc gctcgtgatt tgtttgcaaa 2821 aagaacaaaa ctgaaaaaac ccagacacgc tcgacttcct gtcttcctat tgattgcagc 2881 ttccaatttc gtcacacaac aaggtcctag cgacggctca caggttttgt aacaagcaat 2941 cgaaggttct ggaatggcgg gaaagggttt agtaccacat gctatgatgc ccactgtgat 3001 ctccagagca aagttcgttc gatcgtactg ttactctctc tctttcaaac agaattgtcc 3061 gaatcgtgtg acaacaacag cctgttctca cacactcttt tcttctaacc aagggggtgg 3121 tttagtttag tagaacctcg tgaaacttac atttacatat atataaactt gcataaattg 3181 gtcaatgcaa gaaatacata tttggtcttt tctaattcgt agtttttcaa gttcttagat 3241 gctttctttt tctctttttt acagatcatc aaggaagtaa ttatctactt tttacaacaa 3301 atataaaaca ccggatctca aaatggataa aaagtatagt attggtttag atattggtac 3361 taactctgtg ggttgggcag ttatcactga cgaatataaa gttccatcaa agaaatttaa 3421 ggtgttaggt aacactgaca gacactcaat aaaaaagaat cttatcggtg ctcttttgtt 3481 cgactccggt gaaactgccg aggctacacg tttaaaaaga acagcaagaa gaagatatac 3541 ccgtagaaaa aatagaatat gttatttaca agaaatcttt tctaatgaaa tggctaaagt 3601 tgatgattcc tttttccata gattggaaga gtcatttttg gttgaagaag acaaaaagca 3661 tgagagacat ccaatctttg ggaatatagt tgatgaagtg gcttaccatg aaaaatatcc 3721 taccatttat catttaagaa agaaattggt agattcaact gataaagctg accttagatt 3781 aatctattta gcacttgccc atatgattaa atttagaggt cattttttga ttgaaggtga 3841 tttgaaccca gataattctg acgtggataa attatttatt caattagtcc aaacctacaa 3901 ccaattattt gaggaaaatc caattaatgc tagtggtgtc gatgccaaag ctatattatc 3961 agccagatta tcaaaatcta gacgtttgga aaatttgatt gcccaattgc caggagaaaa 4021 aaagaatgga ttatttggaa acttgatcgc attatcattg ggtttgacac caaattttaa 4081 atctaatttt gatttagctg aagatgctaa attacaatta tcaaaagaca cctatgacga 4141 cgatttggac aatttacttg ctcaaattgg tgatcaatat gcagatttgt tcttagctgc 4201 taaaaactta tctgatgcta ttttgttgtc tgatattttg agagtgaaca cagaaataac 4261 caaagctcca ttatcagcat ctatgatcaa acgttatgat gaacaccatc aggatttgac 4321 tttattgaaa gctttggtga gacaacaatt gccagagaag tataaagaaa tctttttcga 4381 tcaatctaaa aacgggtatg caggttatat tgatgggggt gcctcccaag aggaatttta 4441 caaatttata aaacctattt tagaaaagat ggatgggact gaggaacttt tggtcaaatt 4501 gaacagagaa gatttgttac gtaaacagag aacttttgat aatggtagta tacctcacca 4561 aattcatttg ggtgagttgc atgcaatttt aagaagacaa gaagattttt atccattttt 4621 aaaagataat agagaaaaaa tcgagaaaat tttaaccttt agaattccat actatgttgg 4681 gcctttggct agaggtaatt caagatttgc ctggatgaca cgtaaatcag aagaaactat 4741 taccccttgg aattttgaag aggttgttga taaaggagca tcagcacaga gttttattga 4801 aagaatgacc aatttcgata aaaacttacc aaatgaaaaa gttttaccaa aacattcctt 4861 gttatacgaa tattttactg tttacaatga acttacaaag gttaaatatg ttactgaagg 4921 tatgcgtaag ccagcctttt tatctggaga acagaaaaag gcaatagttg atttattgtt 4981 taaaacaaat agaaaagtta ctgttaaaca attaaaagaa gattacttta agaaaattga 5041 atgttttgat tcagttgaaa tcagtggtgt tgaagacaga tttaatgcta gtttaggaac 5101 ttaccatgat ttacttaaaa ttatcaaaga taaagatttc ttggataacg aagaaaatga 5161 agacatttta gaagacattg ttttaacctt aactttattc gaagatagag agatgattga 5221 agaacgtttg aagacttatg cacatttgtt tgacgataaa gtgatgaaac agttgaaaag 5281 aagacgttat actggatggg gtagattgtc tcgtaaattg atcaatggaa ttagagataa 5341 acaaagtggt aaaactatct tggacttttt gaaatctgac ggatttgcta atagaaattt 5401 catgcaattg atccacgacg atagtttgac atttaaagaa gacatccaaa aggcccaagt 5461 gagtgggcaa ggtgattcat tacatgaaca tattgcaaat ttagccggat ctcctgctat 5521 taagaaaggg atattacaaa ctgttaaagt tgtggatgaa ttagtgaaag taatgggaag 5581 acataaacct gaaaacattg tcattgagat ggcaagagaa aatcaaacta cacaaaaagg 5641 acagaaaaat agtagagaac gtatgaaaag aatagaagag ggtattaaag aattgggtag 5701 tcaaatattg aaagaacacc cagtggaaaa tacccagttg caaaatgaaa aattatatct 5761 ttactacctt caaaatggac gtgatatgta tgttgatcag gaattagata taaatagact 5821 ttcagattat gatgtagatc atatagttcc acaatctttc ttgaaagatg attccataga 5881 caataaagta ttaactagaa gtgataaaaa tagaggtaaa agtgataatg tcccaagtga 5941 ggaagtcgtc aaaaagatga aaaattactg gcgtcaactt ttgaatgcta aattaattac 6001 tcaaagaaaa tttgataatt tgactaaagc agaaagaggt gggctttctg aattagataa 6061 agccgggttc attaaaagac aattggtcga aactagacaa attactaaac atgttgccca 6121 aattttagat tcccgtatga acactaagta tgacgaaaat gataagttaa tacgtgaggt 6181 taaagtcatt actttaaaat caaaacttgt ctctgatttc agaaaggatt tccaattcta 6241 taaagttaga gaaattaata attatcatca tgctcatgat gcatatttga atgctgtagt 6301 tggaactgct ttaatcaaga aataccctaa attagaatct gaatttgtat atggtgatta 6361 caaagtctat gatgttagaa agatgattgc taaatcagaa caagaaattg gtaaagctac 6421 agctaaatac ttcttttact ctaacattat gaatttcttt aaaacagaaa ttactttggc 6481 aaacggtgaa attagaaaaa gacctcttat tgaaacaaat ggtgagactg gagagatagt 6541 ttgggacaaa gggcgtgatt tcgctactgt tagaaaagtt ttatcaatgc cacaagttaa 6601 cattgtaaag aaaacagagg ttcaaactgg tggtttctca aaagaaagta ttttgcctaa 6661 aagaaatagt gataaattga ttgccagaaa aaaggattgg gatccaaaga aatatggtgg 6721 tttcgactca ccaaccgtag cctattctgt tttggttgtg gcaaaggttg aaaagggtaa 6781 aagtaaaaag cttaaatcag taaaagaact tttgggtatt acaataatgg aaagaagttc 6841 ctttgaaaag aaccctattg attttttgga agctaaaggt tataaggaag taaagaagga 6901 cttaataatc aaattgccta aatattcttt atttgaatta gaaaatggga gaaaaagaat 6961 gttggcttct gctggagaat tgcaaaaggg taatgaatta gcattgcctt ccaaatatgt 7021 taacttcttg tatttagctt cacactatga aaagttgaaa gggtcaccag aagataacga 7081 gcaaaaacaa ttatttgttg aacaacacaa acactactta gatgagatta tagaacaaat 7141 tagtgaattc agtaaaagag tgatattagc tgatgcaaat ttagataaag ttttgtcagc 7201 ctataacaaa catagagata agccaattag agaacaagca gaaaacatta ttcacttatt 7261 tacccttacc aatttaggag cacctgctgc tttcaagtat tttgatacaa caattgatcg 7321 taaaagatat acctcaacaa aagaagtctt agacgccacc ttaattcatc aatcaatcac 7381 tggattgtat gagacaagaa ttgatttgtc tcaattgggt ggtgatgaag gggctgatcc 7441 taagaagaaa agaaaagttg atccaaagaa aaagcgtaag gtggatccta agaaaaagag 7501 aaaggttgac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 7561 tgacgatgac aagtgataat ctcttctcga gtcatgtaat tagttatgtc acgcttacat 7621 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc 7681 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa 7741 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg 7801 cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtactg cggtgtgaaa 7861 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt 7921 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 7981 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 8041 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 8101 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 8161 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 8221 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 8281 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 8341 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 8401 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt 8461 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa 8521 tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt 8581 ggatcccccg ggctgcagga attcgatatc aagctttctt tgaaaagata atgtatgatt 8641 atgctttcac tcatatttat acagaaactt gatgttttct ttcgagtata tacaaggtga 8701 ttacatgtac gtttgaagta caactctaga ttttgtagtg ccctcttggg ctagcggtaa 8761 aggtgcgcat tttttcacac cctacaatgt tctgttcaaa agattttggt caaacgctgt 8821 agaagtgaaa gttggtgcgc atgtttcggc gttcgaaact tctccgcagt gaaagataaa 8881 tgatcctaga tgcgagatat cctcatgcct aggttttaga gctagaaata gcaagttaaa 8941 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtggtg ctttttttgt 9001 tttttatgtc taccgtcgac ctcgaggggg ggcccggtac cagcttttgt tccctttagt 9061 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 9121 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 9181 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 9241 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 9301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 9361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 9421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 9481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 9541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 9601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 9661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 9721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 9781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 9841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 9901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 9961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 10021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 10081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 10141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 10201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 10261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 10321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 10381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 10441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 10501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 10561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 10621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 10681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 10741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 10801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 10861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 10921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 10981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 11041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 11101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 11161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 11221 catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa 11281 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag 11341 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac 11401 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct 11461 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat 11521 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg 11581 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc 11641 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt 11701 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg 11761 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct 11821 ttcgtc //