LOCUS Exported 10073 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION D435C/E945C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, lobe closure FRET construct. ACCESSION . VERSION . KEYWORDS pSHS293 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10073) AUTHORS Sternberg SH, LaFrance B, Kaplan M, Doudna JA TITLE Conformational control of DNA target cleavage by CRISPR-Cas9. JOURNAL Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28. PUBMED 26524520 REFERENCE 2 (bases 1 to 10073) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10073 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 213..230 /label=L4440 /note="L4440 vector, forward primer" misc_feature 245..384 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(310..329) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 470..492 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(486..677) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 2135..2154 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 2238..2257 /label=T7 /note="T7 promoter, forward primer" promoter 2238..2256 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 2345..2367 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2387..2416 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 2426..3526 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" primer_bind 3500..3523 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 3581..3601 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 3608..7714 /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 NLIGALLFDSGETAEATRLKRTARRRYTRRKNRISYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKCNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDCNDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" primer_bind complement(7840..7858) /label=T7 Term /note="T7 terminator, reverse primer" terminator 7854..7901 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(8267..8295) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(8322..8340) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8408..8512 /gene="bla" /label=AmpR promoter CDS 8513..9373 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSR?DAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMP?AMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8731..8750) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9544..59 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10033..10052 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 61 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 121 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 181 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 241 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg 301 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg 361 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga 421 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 481 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca 541 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg 601 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt 661 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg 721 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg 781 ttactggaac gtcgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa 841 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc 901 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt 961 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac 1021 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca 1081 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc 1141 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac 1201 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga 1261 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg 1321 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg 1381 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg 1441 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa 1501 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc 1561 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga 1621 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct 1681 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc 1741 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga 1801 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca 1861 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg 1921 ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca 1981 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg 2041 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca 2101 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag 2161 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga 2221 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag 2281 tgccggctcc ggagagctct ttaattaagc ggccgccctg caggactcga gttctagaaa 2341 taattttgtt taactttaag aaggagatat acatatgaaa tcttctcacc atcaccatca 2401 ccatcaccat caccatggtt cttctatgaa aatcgaagaa ggtaaactgg taatctggat 2461 taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac 2521 cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc 2581 ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc 2641 tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc 2701 gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga 2761 agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga 2821 gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct 2881 gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta 2941 tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg caaaagcggg 3001 tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc 3061 catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc 3121 atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa 3181 gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc 3241 gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga 3301 agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt 3361 ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc cagaaaggtg aaatcatgcc 3421 gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc 3481 cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactaatt cgagctcgaa 3541 caacaacaac aataacaata acaacaacct cgggatcgag gaaaacctgt acttccaatc 3601 caatgcaatg gataagaaat actcaatagg cttagatatc ggcacaaata gcgtcggatg 3661 ggcggtgatc actgatgaat ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac 3721 agaccgccac agtatcaaaa aaaatcttat aggggctctt ttatttgaca gtggagagac 3781 agcggaagcg actcgtctca aacggacagc tcgtagaagg tatacacgtc ggaagaatcg 3841 tatttcttat ctacaggaga ttttttcaaa tgagatggcg aaagtagatg atagtttctt 3901 tcatcgactt gaagagtctt ttttggtgga agaagacaag aagcatgaac gtcatcctat 3961 ttttggaaat atagtagatg aagttgctta tcatgagaaa tatccaacta tctatcatct 4021 gcgaaaaaaa ttggtagatt ctactgataa agcggatttg cgcttaatct atttggcctt 4081 agcgcatatg attaagtttc gtggtcattt tttgattgag ggagatttaa atcctgataa 4141 tagtgatgtg gacaaactat ttatccagtt ggtacaaacc tacaatcaat tatttgaaga 4201 aaaccctatt aacgcaagtg gagtagatgc taaagcgatt ctttctgcac gattgagtaa 4261 atcaagacga ttagaaaatc tcattgctca gctccccggt gagaagaaaa atggcttatt 4321 tgggaatctc attgctttgt cattgggttt gacccctaat tttaaatcaa attttgattt 4381 ggcagaagat gctaaattac agctttcaaa agatacttac gatgatgatt tagataattt 4441 attggcgcaa attggagatc aatatgctga tttgtttttg gcagctaaga atttatcaga 4501 tgctatttta ctttcagata tcctaagagt aaatactgaa ataactaagg ctcccctatc 4561 agcttcaatg attaaacgct acgatgaaca tcatcaagac ttgactcttt taaaagcttt 4621 agttcgacaa caacttccag aaaagtataa agaaatcttt tttgatcaat caaaaaacgg 4681 atatgcaggt tatattgatg ggggagctag ccaagaagaa ttttataaat ttatcaaacc 4741 aattttagaa aaaatggatg gtactgagga attattggtg aaactaaatc gtgaagattt 4801 gctgcgcaag caacggacct ttgacaacgg ctctattccc catcaaattc acttgggtga 4861 gctgcatgct attttgagaa gacaagaaga cttttatcca tttttaaaat gcaatcgtga 4921 gaagattgaa aaaatcttga cttttcgaat tccttattat gttggtccat tggcgcgtgg 4981 caatagtcgt tttgcatgga tgactcggaa gtctgaagaa acaattaccc catggaattt 5041 tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt attgaacgca tgacaaactt 5101 tgataaaaat cttccaaatg aaaaagtact accaaaacat agtttgcttt atgagtattt 5161 tacggtttat aacgaattga caaaggtcaa atatgttact gaaggaatgc gaaaaccagc 5221 atttctttca ggtgaacaga agaaagccat tgttgattta ctcttcaaaa caaatcgaaa 5281 agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa atagaatctt ttgatagtgt 5341 tgaaatttca ggagttgaag atagatttaa tgcttcatta ggtacctacc atgatttgct 5401 aaaaattatt aaagataaag attttttgga taatgaagaa aatgaagata tcttagagga 5461 tattgtttta acattgacct tatttgaaga tagggagatg attgaggaaa gacttaaaac 5521 atatgctcac ctctttgatg ataaggtgat gaaacagctt aaacgtcgcc gttatactgg 5581 ttggggacgt ttgtctcgaa aattgattaa tggtattagg gataagcaat ctggcaaaac 5641 aatattagat tttttgaaat cagatggttt tgccaatcgc aattttatgc agctgatcca 5701 tgatgatagt ttgacattta aagaagacat tcaaaaagca caagtgtctg gacaaggcga 5761 tagtttacat gaacatattg caaatttagc tggtagccct gctattaaaa aaggtatttt 5821 acagactgta aaagttgttg atgaattggt caaagtaatg gggcggcata agccagaaaa 5881 tatcgttatt gaaatggcac gtgaaaatca gacaactcaa aagggccaga aaaattcgcg 5941 agagcgtatg aaacgaatcg aagaaggtat caaagaatta ggaagtcaga ttcttaaaga 6001 gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc tatctctatt atctccaaaa 6061 tggaagagac atgtatgtgg accaagaatt agatattaat cgtttaagtg attatgatgt 6121 cgatcacatt gttccacaaa gtttccttaa agacgattca atagacaata aggtcttaac 6181 gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa 6241 gatgaaaaac tattggagac aacttctaaa cgccaagtta atcactcaac gtaagtttga 6301 taatttaacg aaagctgaac gtggaggttt gagtgaactt gataaagctg gttttatcaa 6361 acgccaattg gttgaaactc gccaaatcac taagcatgtg gcacaaattt tggatagtcg 6421 catgaatact aaatacgatt gcaatgataa acttattcga gaggttaaag tgattacctt 6481 aaaatctaaa ttagtttctg acttccgaaa agatttccaa ttctataaag tacgtgagat 6541 taacaattac catcatgccc atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat 6601 taagaaatat ccaaaacttg aatcggagtt tgtctatggt gattataaag tttatgatgt 6661 tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt 6721 ttactctaat atcatgaact tcttcaaaac agaaattaca cttgcaaatg gagagattcg 6781 caaacgccct ctaatcgaaa ctaatgggga aactggagaa attgtctggg ataaagggcg 6841 agattttgcc acagtgcgca aagtattgtc catgccccaa gtcaatattg tcaagaaaac 6901 agaagtacag acaggcggat tctccaagga gtcaatttta ccaaaaagaa attcggacaa 6961 gcttattgct cgtaaaaaag actgggatcc aaaaaaatat ggtggttttg atagtccaac 7021 ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa gggaaatcga agaagttaaa 7081 atccgttaaa gagttactag ggatcacaat tatggaaaga agttcctttg aaaaaaatcc 7141 gattgacttt ttagaagcta aaggatataa ggaagttaaa aaagacttaa tcattaaact 7201 acctaaatat agtctttttg agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg 7261 agaattacaa aaaggaaatg agctggctct gccaagcaaa tatgtgaatt ttttatattt 7321 agctagtcat tatgaaaagt tgaagggtag tccagaagat aacgaacaaa aacaattgtt 7381 tgtggagcag cataagcatt atttagatga gattattgag caaatcagtg aattttctaa 7441 gcgtgttatt ttagcagatg ccaatttaga taaagttctt agtgcatata acaaacatag 7501 agacaaacca atacgtgaac aagcagaaaa tattattcat ttatttacgt tgacgaatct 7561 tggagctccc gctgctttta aatattttga tacaacaatt gatcgtaaac gatatacgtc 7621 tacaaaagaa gttttagatg ccactcttat ccatcaatcc atcactggtc tttatgaaac 7681 acgcattgat ttgagtcagc taggaggtga ctaataacat tggaagtgga taacggatcc 7741 gcgatcgcgg cgcgccacct ggtggccggc cggtaccacg cgtgcgcgct gatccggctg 7801 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat 7861 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat 7921 ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt 7981 agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt 8041 gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg 8101 ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg 8161 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata 8221 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg 8281 atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt 8341 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 8401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 8461 catgagacaa taaccctgat aaatgcttca ataacattga aaaaggaaga gtatgagtat 8521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 8581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 8641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 8701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtrttga 8761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 8821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 8881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 8941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 9001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgyagc 9061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 9121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 9181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 9241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 9301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 9361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 9421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 9481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 9541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 9601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 9661 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 9721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 9781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 9841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 9901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 9961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 10021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gcc //