LOCUS Exported 10073 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION S867C/S355C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, HNH-1 FRET construct. ACCESSION . VERSION . KEYWORDS pSHS306 - Bacterial expression plasmid for SpCas9, HNH FRET va... 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 63..82 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 166..185 /label=T7 /note="T7 promoter, forward primer" promoter 166..184 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 273..295 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 315..344 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 354..1454 /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 1428..1451 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 1509..1529 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1536..5642 /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 QQLPEKYKEIFFDQCKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKCDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" primer_bind complement(5768..5786) /label=T7 Term /note="T7 terminator, reverse primer" terminator 5782..5829 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(6195..6223) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(6250..6268) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6336..6440 /gene="bla" /label=AmpR promoter CDS 6441..7301 /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(6659..6678) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7472..8060 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7961..7980 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8214..8231 /label=L4440 /note="L4440 vector, forward primer" misc_feature 8246..8385 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(8311..8330) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8471..8493 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(8487..8678) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca 61 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc 121 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta 181 tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt aattaagcgg 241 ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa ggagatatac 301 atatgaaatc ttctcaccat caccatcacc atcaccatca ccatggttct tctatgaaaa 361 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg 421 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata 481 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct 541 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg 601 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca 661 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc 721 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga 781 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg 841 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg 901 gcgtggataa cgctggcgca aaagcgggtc tgaccttcct ggttgacctg attaaaaaca 961 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa 1021 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt 1081 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc 1141 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa 1201 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg 1261 tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccactatgg 1321 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg 1381 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga 1441 aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac aacaacctcg 1501 ggatcgagga aaacctgtac ttccaatcca atgcaatgga taagaaatac tcaataggct 1561 tagatatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt 1621 ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag 1681 gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa cggacagctc 1741 gtagaaggta tacacgtcgg aagaatcgta tttcttatct acaggagatt ttttcaaatg 1801 agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag 1861 aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc 1921 atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag 1981 cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt 2041 tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg 2101 tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta 2161 aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc 2221 tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga 2281 cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag 2341 atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt 2401 tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa 2461 atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc 2521 atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag 2581 aaatcttttt tgatcaatgt aaaaacggat atgcaggtta tattgatggg ggagctagcc 2641 aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat 2701 tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct 2761 ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact 2821 tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgaattc 2881 cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt 2941 ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc 3001 aatcatttat tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac 3061 caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat 3121 atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg 3181 ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt 3241 tcaaaaaaat agaatctttt gatagtgttg aaatttcagg agttgaagat agatttaatg 3301 cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat tttttggata 3361 atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata 3421 gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga 3481 aacagcttaa acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg 3541 gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg 3601 ccaatcgcaa ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc 3661 aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg 3721 gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca 3781 aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga 3841 caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca 3901 aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg 3961 aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag 4021 atattaatcg tttaagtgat tatgatgtcg atcacattgt tccacaaagt ttccttaaag 4081 acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt aaatgcgata 4141 acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg 4201 ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga 4261 gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta 4321 agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac 4381 ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag 4441 atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc 4501 taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg 4561 tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa 4621 taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag 4681 aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa 4741 ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca 4801 tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt 4861 caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa 4921 aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg 4981 tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta 5041 tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg 5101 aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg 5161 gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc 5221 caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc 5281 cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga 5341 ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata 5401 aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata 5461 ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata 5521 caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc 5581 atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgact 5641 aataacattg gaagtggata acggatccgc gatcgcggcg cgccacctgg tggccggccg 5701 gtaccacgcg tgcgctctga tccggctgct aacaaagccc gaaaggaagc tgagttggct 5761 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg 5821 ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat 5881 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa 5941 gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc 6001 tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc 6061 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat 6121 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg 6181 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa 6241 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 6301 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 6361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 6421 aacattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 6481 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 6541 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 6601 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 6661 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac 6721 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 6781 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 6841 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 6901 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 6961 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg 7021 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 7081 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 7141 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 7201 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 7261 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 7321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 7381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 7441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 7501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 7561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc 7621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 7681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 7741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 7801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 7861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 7921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 7981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 8041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 8101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 8161 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 8221 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg 8281 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag 8341 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca 8401 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct 8461 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg 8521 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt 8581 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag 8641 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga 8701 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt 8761 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg 8821 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat 8881 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg 8941 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat 9001 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc 9061 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac 9121 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg 9181 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc 9241 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg 9301 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg 9361 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg 9421 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa 9481 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca 9541 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg 9601 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc 9661 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt 9721 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc 9781 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt 9841 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga 9901 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact 9961 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga 10021 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gcc //