LOCUS Exported 9075 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express Streptococcus pyogenes Cas9. ACCESSION . VERSION . KEYWORDS SpyCas9(WT) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9075) AUTHORS Dong, Guo M, Wang S, Zhu Y, Wang S, Xiong Z, Yang J, Xu Z, Huang Z TITLE Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. JOURNAL Nature. 2017 Jun 15;546(7658):436-439. doi: 10.1038/nature22377. Epub 2017 Apr 27. PUBMED 28448066 REFERENCE 2 (bases 1 to 9075) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9075 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 530..560 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 568..584 /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 592..608 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 592..608 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 604..1131 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(624..641) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(624..640) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(633..655) /label=M13/pUC Forward /note="In lacZ gene" promoter 1233..1261 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 1269..1285 /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)." CDS 1308..1961 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 1919..1941 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 1968..1991 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 2001..6104 /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" primer_bind complement(6175..6197) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6342..6360) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6428..6532 /gene="bla" /label=AmpR promoter CDS 6533..7393 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6751..6770) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7564..8152 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8053..8072 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8306..8323 /label=L4440 /note="L4440 vector, forward primer" promoter 8396..8473 /gene="lacI" /label=lacI promoter CDS 8474..481 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(8493..8512) /label=LacI-R /note="LacI, reverse primer" ORIGIN 1 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag 61 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc 121 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg 181 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg 241 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt 301 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt 361 ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct ctccccgcgc 421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 481 agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct ttacacttta 541 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 601 gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac 661 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat 721 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg 781 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt 841 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc 901 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat 961 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag 1021 acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct 1081 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca 1141 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat 1201 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat 1261 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg tcccctatac 1321 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg 1381 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa 1441 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat 1501 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt 1561 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg 1621 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca 1681 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg 1741 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca 1801 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag 1861 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg 1921 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gaagttctgt 1981 tccaggggcc cctgggatcc atggacaaga agtacagcat cggcctggac atcggcacca 2041 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg 2101 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg 2161 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca 2221 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg 2281 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg 2341 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca 2401 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga 2461 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc 2521 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc 2581 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg 2641 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga 2701 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga 2761 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg 2821 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca 2881 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca 2941 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc 3001 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc 3061 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca 3121 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga 3181 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga 3241 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga 3301 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc 3361 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca 3421 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc 3481 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc 3541 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa 3601 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca 3661 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt 3721 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat 3781 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg 3841 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg 3901 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc 3961 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc 4021 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca 4081 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt 4141 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta 4201 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc 4261 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac 4321 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc 4381 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt 4441 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt 4501 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca 4561 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag 4621 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc 4681 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg 4741 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga 4801 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga 4861 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca 4921 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg 4981 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca 5041 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg 5101 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca 5161 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt 5221 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata 5281 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga 5341 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct 5401 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt 5461 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct 5521 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc 5581 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc 5641 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga 5701 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc 5761 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca 5821 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct 5881 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta 5941 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga 6001 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg 6061 gcctgtacga gacacggatc gacctgtctc agctgggagg cgatgatgac tcgagcggcc 6121 gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg 6181 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 6241 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc 6301 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt 6361 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 6421 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 6481 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 6541 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 6601 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 6661 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 6721 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga 6781 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 6841 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 6901 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 6961 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 7021 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc 7081 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 7141 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 7201 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 7261 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 7321 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 7381 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 7441 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 7501 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 7561 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 7621 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 7681 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 7741 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 7801 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 7861 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 7921 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 7981 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 8041 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 8101 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 8161 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 8221 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 8281 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct 8341 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac 8401 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt 8461 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc 8521 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga 8581 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact 8641 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc 8701 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt 8761 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc 8821 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt 8881 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat 8941 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc 9001 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct 9061 gcgtctggct ggctg //