LOCUS Exported 10610 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR-Cas9 system for genetic manipulation of Candida tropicalis. ACCESSION . VERSION . KEYWORDS pCT-tRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10610) AUTHORS Lombardi L, Oliveira-Pacheco J, Butler G TITLE Plasmid-Based CRISPR-Cas9 Gene Editing in Multiple Candida Species. JOURNAL mSphere. 2019 Mar 13;4(2). pii: 4/2/e00125-19. doi: 10.1128/mSphere.00125-19. PUBMED 30867327 REFERENCE 2 (bases 1 to 10610) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10610 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..344 /label=TEF promoter /note="Ashbya gossypii TEF promoter" misc_RNA 458..533 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(629..647) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" terminator 659..848 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind 995..1017 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1009..1026 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1010..1026 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1033..1052 /label=T7 /note="T7 promoter, forward primer" promoter 1033..1051 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3194..7297 /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 7310..7330 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7334..7354 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7358..7378 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7379..7444 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" promoter complement(7679..7697) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(7680..7697) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(7715..7731) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7715..7731) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7728..7750) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7739..7755 /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(7763..7793) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7808..7829 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(8767..9355) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8847..8866) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9526..10386) /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 10149..10168 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10387..10491) /gene="bla" /label=AmpR promoter ORIGIN 1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga 61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata 121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgcg gacctgcgag 181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga 241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc 301 ttgctaggat acagttctca catcacatcc gaacataaac aaccaaacaa agagcttaaa 361 atgggcgtgt ggcgtagttg gtagcgcgtt cccttagcat gggaaaggtc atgagttcga 421 ttcttatctc gtccaggaag agctcgcgag ctcttccgtt ttagagctag aaatagcaag 481 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttggc 541 cggcatggtc ccagcctcct cgctggcgcc ggctgggcaa catgcttcgg catggcgaat 601 gggactctcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat 661 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt 721 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt 781 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac 841 gctcgaaggc tttaacgagc tcactagttc cattcgccat tcaggctgcg caactgttgg 901 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct 961 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg 1021 gccagtgaat tgtaatacga ctcactatag ggcgaattgg gcccgtccaa ctggtggatc 1081 agtatcggtt atgtcatcta aaaacaaaat tgatgctgca aaacgatgga atatcccagt 1141 tgtctcggtt gcgtatttgt gggaaatttt agaactctcg actaacaaat cgcatcttat 1201 catgcccgat attacagatt tacagtggtg tatttttgca cctagcaact ataataaacc 1261 taaatcatta ttggagtatg taaagaattt ggataaagcc agtagagaaa gttcctttgg 1321 tcccaagagt caagaaattg atgcacaaga acaacctaca atggataatc tggtgaagtt 1381 gccctcacca cgaagagtta acctgaaaca aaaatatggt aaattggtac gaggtaaatc 1441 tccgaaatca attaaacgga aattactaga agctgcaaat tcgtttactg atggtcaaaa 1501 tgatcacgat attaatccag atgtaacact tgaagaggac atgatgtcac aaataagata 1561 tcaagacaat gagtctatga tcaaccaaga aagattatta gagaaattgg atggatcagc 1621 tttaccactg gagaccctga tacagcatac ctgtaaacgg agtagaaaaa aataactgta 1681 taatatttgc tattactata aaacttgtat cattattctt atatggggtt gtagccctaa 1741 ttactcttac tttctcgtga tttgcacgtg tttgtttttg tatttgtgcg actaatatta 1801 tagcgcattt gttatcttag tgaaatcagt catacctttt gttctccccc taactatcta 1861 ctattctcat gaaactgcaa gaaaaaagtt ctgctgttca ctttctctcg ttcaactttt 1921 acaaagtaaa aaaaaaaaaa aaaatttttt ttttcctcgt catcttgatt ttacttcttc 1981 ttctttttcc tttttccttt cttcacactt tatagtcata atcaatcatc gattgctaga 2041 attaccggat ctatgaaaat ttcggtgatc cctgagcagg tggcggaaac attggatgct 2101 gagaaccatt tcattgttcg tgaagtgttc gatgtgcacc tatccgacca aggctttgaa 2161 ctatctacca gaagtgtgag cccctaccgg aaggattaca tctcggatga tgactctgat 2221 gaagactctg cttgctatgg cgcattcatc gaccaagagc ttgtcgggaa gattgaactc 2281 aactcaacat ggaacgatct agcctctatc gaacacattg ttgtgtcgca cacgcaccga 2341 ggcaaaggag tcgcgcacag tctcatcgaa tttgcgaaaa agtgggcact aagcagacag 2401 ctccttggca tacgattaga gacacaaacg aacaatgtac ctgcctgcaa tttgtacgca 2461 aaatgtggct ttactctcgg cggcattgac ctcttcacgt ataaaactag acctcaagtc 2521 tcgaacgaaa cagcgatgta ctggtactgg ttctcgggag cacaggatga cgccatgcag 2581 ggcgcctaag aatatacgtc gttattttgt gttatacact cagctacaat ttcgatttcc 2641 cgctcatgat gaggccccat gcttcctctg tcaactcctc gtacttgtca ccattcaacc 2701 agtagatcac atcggtgccg ctttctccct tgggaagttt ttggttctta aattggttct 2761 tgggcacctt gtggttatgt gagcggccgc ggtaccgacg tcacccgctc tgacggttac 2821 ccaatgcggt tataagccaa cagtctgttg tgcgactagg ctcgcttggc acctgcacag 2881 atgctgcgac agctctcacg cacagaaatg gtcacctaga gtcgatttcc gcgcctcgtt 2941 gccgccggtc tccgcgcggt gaatcctgta catagtcatc tccgattcac tttcactaga 3001 cgaatccggc acatgagtga tccggcgtgc acacaatagc aatctccctg cacacaccgg 3061 gacgcgattg ccgggtaatc cctggttggg tcgtttctgc ctcgttgttt gataccagcg 3121 ctcacccctt tcgaaaaatt tacttttgac taggtattaa tatagtatag caaactgcag 3181 ctcgagggat ccgatggata aaaagtatag tattggttta gatattggta ctaactctgt 3241 gggttgggca gttatcactg acgaatataa agttccatca aagaaattta aggtgttagg 3301 taacactgac agacactcaa taaaaaagaa tcttatcggt gctcttttgt tcgactccgg 3361 tgaaactgcc gaggctacac gtttaaaaag aacagcaaga agaagatata cccgtagaaa 3421 aaatagaata tgttatttac aagaaatctt ttctaatgaa atggctaaag ttgatgattc 3481 ctttttccat agattggaag agtcattttt ggttgaagaa gacaaaaagc atgagagaca 3541 tccaatcttt gggaatatag ttgatgaagt ggcttaccat gaaaaatatc ctaccattta 3601 tcatttaaga aagaaattgg tagattcaac tgataaagct gaccttagat taatctattt 3661 agcacttgcc catatgatta aatttagagg tcattttttg attgaaggtg atttgaaccc 3721 agataattct gacgtggata aattatttat tcaattagtc caaacctaca accaattatt 3781 tgaggaaaat ccaattaatg ctagtggtgt cgatgccaaa gctatattat cagccagatt 3841 atcaaaatct agacgtttgg aaaatttgat tgcccaattg ccaggagaaa aaaagaatgg 3901 attatttgga aacttgatcg cattatcatt gggtttgaca ccaaatttta aatctaattt 3961 tgatttagct gaagatgcta aattacaatt atcaaaagac acctatgacg acgatttgga 4021 caatttactt gctcaaattg gtgatcaata tgcagatttg ttcttagctg ctaaaaactt 4081 atctgatgct attttgttgt ctgatatttt gagagtgaac acagaaataa ccaaagctcc 4141 attatcagca tctatgatca aacgttatga tgaacaccat caggatttga ctttattgaa 4201 agctttggtg agacaacaat tgccagagaa gtataaagaa atctttttcg atcaatctaa 4261 aaacgggtat gcaggttata ttgatggggg tgcctcccaa gaggaatttt acaaatttat 4321 aaaacctatt ttagaaaaga tggatgggac tgaggaactt ttggtcaaat tgaacagaga 4381 agatttgtta cgtaaacaga gaacttttga taatggtagt atacctcacc aaattcattt 4441 gggtgagttg catgcaattt taagaagaca agaagatttt tatccatttt taaaagataa 4501 tagagaaaaa atcgagaaaa ttttaacctt tagaattcca tactatgttg ggcctttggc 4561 tagaggtaat tcaagatttg cctggatgac acgtaaatca gaagaaacta ttaccccttg 4621 gaattttgaa gaggttgttg ataaaggagc atcagcacag agttttattg aaagaatgac 4681 caatttcgat aaaaacttac caaatgaaaa agttttacca aaacattcct tgttatacga 4741 atattttact gtttacaatg aacttacaaa ggttaaatat gttactgaag gtatgcgtaa 4801 gccagccttt ttatctggag aacagaaaaa ggcaatagtt gatttattgt ttaaaacaaa 4861 tagaaaagtt actgttaaac aattaaaaga agattacttt aagaaaattg aatgttttga 4921 ttcagttgaa atcagtggtg ttgaagacag atttaatgct agtttaggaa cttaccatga 4981 tttacttaaa attatcaaag ataaagattt cttggataac gaagaaaatg aagacatttt 5041 agaagacatt gttttaacct taactttatt cgaagataga gagatgattg aagaacgttt 5101 gaagacttat gcacatttgt ttgacgataa agtgatgaaa cagttgaaaa gaagacgtta 5161 tactggatgg ggtagattgt ctcgtaaatt gatcaatgga attagagata aacaaagtgg 5221 taaaactatc ttggactttt tgaaatctga cggatttgct aatagaaatt tcatgcaatt 5281 gatccacgac gatagtttga catttaaaga agacatccaa aaggcccaag tgagtgggca 5341 aggtgattca ttacatgaac atattgcaaa tttagccgga tctcctgcta ttaagaaagg 5401 gatattacaa actgttaaag ttgtggatga attagtgaaa gtaatgggaa gacataaacc 5461 tgaaaacatt gtcattgaga tggcaagaga aaatcaaact acacaaaaag gacagaaaaa 5521 tagtagagaa cgtatgaaaa gaatagaaga gggtattaaa gaattgggta gtcaaatatt 5581 gaaagaacac ccagtggaaa atacccagtt gcaaaatgaa aaattatatc tttactacct 5641 tcaaaatgga cgtgatatgt atgttgatca ggaattagat ataaatagac tttcagatta 5701 tgatgtagat catatagttc cacaatcttt cttgaaagat gattccatag acaataaagt 5761 attaactaga agtgataaaa atagaggtaa aagtgataat gtcccaagtg aggaagtcgt 5821 caaaaagatg aaaaattact ggcgtcaact tttgaatgct aaattaatta ctcaaagaaa 5881 atttgataat ttgactaaag cagaaagagg tgggctttct gaattagata aagccgggtt 5941 cattaaaaga caattggtcg aaactagaca aattactaaa catgttgccc aaattttaga 6001 ttcccgtatg aacactaagt atgacgaaaa tgataagtta atacgtgagg ttaaagtcat 6061 tactttaaaa tcaaaacttg tctctgattt cagaaaggat ttccaattct ataaagttag 6121 agaaattaat aattatcatc atgctcatga tgcatatttg aatgctgtag ttggaactgc 6181 tttaatcaag aaatacccta aattagaatc tgaatttgta tatggtgatt acaaagtcta 6241 tgatgttaga aagatgattg ctaaatcaga acaagaaatt ggtaaagcta cagctaaata 6301 cttcttttac tctaacatta tgaatttctt taaaacagaa attactttgg caaacggtga 6361 aattagaaaa agacctctta ttgaaacaaa tggtgagact ggagagatag tttgggacaa 6421 agggcgtgat ttcgctactg ttagaaaagt tttatcaatg ccacaagtta acattgtaaa 6481 gaaaacagag gttcaaactg gtggtttctc aaaagaaagt attttgccta aaagaaatag 6541 tgataaattg attgccagaa aaaaggattg ggatccaaag aaatatggtg gtttcgactc 6601 accaaccgta gcctattctg ttttggttgt ggcaaaggtt gaaaagggta aaagtaaaaa 6661 gcttaaatca gtaaaagaac ttttgggtat tacaataatg gaaagaagtt cctttgaaaa 6721 gaaccctatt gattttttgg aagctaaagg ttataaggaa gtaaagaagg acttaataat 6781 caaattgcct aaatattctt tatttgaatt agaaaatggg agaaaaagaa tgttggcttc 6841 tgctggagaa ttgcaaaagg gtaatgaatt agcattgcct tccaaatatg ttaacttctt 6901 gtatttagct tcacactatg aaaagttgaa agggtcacca gaagataacg agcaaaaaca 6961 attatttgtt gaacaacaca aacactactt agatgagatt atagaacaaa ttagtgaatt 7021 cagtaaaaga gtgatattag ctgatgcaaa tttagataaa gttttgtcag cctataacaa 7081 acatagagat aagccaatta gagaacaagc agaaaacatt attcacttat ttacccttac 7141 caatttagga gcacctgctg ctttcaagta ttttgataca acaattgatc gtaaaagata 7201 tacctcaaca aaagaagtct tagacgccac cttaattcat caatcaatca ctggattgta 7261 tgagacaaga attgatttgt ctcaattggg tggtgatgaa ggggctgatc ctaagaagaa 7321 aagaaaagtt gatccaaaga aaaagcgtaa ggtggatcct aagaaaaaga gaaaggttga 7381 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 7441 caagtgatat ctcctaggta acaaagcata caagtgtaag aatttcaatt ttatcagtaa 7501 cttattttac cttataatat atgtttatta ctcctacttc tctagtcttc ttggtcccaa 7561 tttccaggtc tacgttgctg aggaacttgg ggaccaccag cacgcttggc catgatgatt 7621 tggtcgatgg acaagatggt ggtagcggct tcagtagcca aatgcatagc ttgagtattc 7681 tatagtgtca cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 7741 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 7801 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 7861 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 7921 tgcgtattga aaaatccggg gtagcgatga ggtagtgcaa gttataccag agagcctcaa 7981 tttgaacgtg gtactgcaca caaaccgtag ccctaaccct aattaaatgc acgtgaccca 8041 cacaattttc aaccaccaac aacacaccaa aaatgtatgt acactcggag tggaaaatat 8101 ttccccaggc ataattgtgg ttgccaatta tagtggagtg tttgttatta gtataggagt 8161 actcctttta tgtggaaaaa tcgaaaaaca taaaacagtg cagctataca ctaacaaatg 8221 gtatacccat ggtcagacaa caccaaacaa cacacccgta atggttggtt tggctaaata 8281 gagcatctca ttttgtgtat tattttatgt tttggatatt tttagcttga aatttatata 8341 aaaatatttt actccaattt tcttgccaaa ttttgtacaa aaagtaaaaa atagaacttc 8401 caatttttgt taaacaaact tcaatctaac aaccgggata gtgtttgggg ggctgtgtaa 8461 caccactggt gtaataaaag tgccaatatt tggaaattaa ttttttggtg gtggacgttt 8521 cttcctaaaa cacctagatg tgctagtata attgcacaac cagcacgcac actgcgtctg 8581 gccggtcctg gactacattt tgtctctaac attcgtagct gcagttcggc tgcggcgagc 8641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 8701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 8761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 8821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 8881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 8941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 9001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 9061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 9121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 9181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 9241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 9301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 9361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 9421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 9481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 9541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 9601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 9661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 9721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 9781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 9841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 9901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 9961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 10021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 10081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 10141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 10201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 10261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 10321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 10381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 10441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 10501 ccgaaaagtg ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 10561 gcatcaggaa ccggtagctt gcctcgtccc cgccgggtca cccggccagc //