LOCUS Exported 10844 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For expression of modified SpyCas9 in e.coli.. ACCESSION . VERSION . KEYWORDS pMJ915v2 + sfGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10844) AUTHORS Staahl BT, Benekareddy M, Coulon-Bainier C, Banfal AA, Floor SN, Sabo JK, Urnes C, Munares GA, Ghosh A, Doudna JA TITLE Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. JOURNAL Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. PUBMED 28191903 REFERENCE 2 (bases 1 to 10844) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10844 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1018..1041 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 1099..1119 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1135..5238 /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 5245..5265 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5272..5292 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(6141..6159) /label=T7 Term /note="T7 terminator, reverse primer" terminator 6155..6202 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(6568..6596) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(6623..6641) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6709..6813 /gene="bla" /label=AmpR promoter CDS 6814..7674 /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(7032..7051) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7845..8433 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8334..8353 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8587..8604 /label=L4440 /note="L4440 vector, forward primer" misc_feature 8619..8758 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(8684..8703) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8844..8866 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(8860..9051) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 10509..10528 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 10612..10631 /label=T7 /note="T7 promoter, forward primer" promoter 10612..10630 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 10719..10741 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 10761..10778 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 10788..1044 /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" ORIGIN 1 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag 61 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac 121 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa 181 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt 241 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac 301 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa 361 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg 421 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt 481 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg 541 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat 601 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc 661 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc 721 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag 781 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg 841 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc 901 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct 961 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat 1021 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac 1081 aacaacctcg ggatcgagga aaacctgtac ttccaatcca atgccactag taccatggat 1141 aagaaatact caataggctt agatatcggc acaaatagcg tcggatgggc ggtgatcact 1201 gatgaatata aggttccgtc taaaaagttc aaggttctgg gaaatacaga ccgccacagt 1261 atcaaaaaaa atcttatagg ggctctttta tttgacagtg gagagacagc ggaagcgact 1321 cgtctcaaac ggacagctcg tagaaggtat acacgtcgga agaatcgtat ttgttatcta 1381 caggagattt tttcaaatga gatggcgaaa gtagatgata gtttctttca tcgacttgaa 1441 gagtcttttt tggtggaaga agacaagaag catgaacgtc atcctatttt tggaaatata 1501 gtagatgaag ttgcttatca tgagaaatat ccaactatct atcatctgcg aaaaaaattg 1561 gtagattcta ctgataaagc ggatttgcgc ttaatctatt tggccttagc gcatatgatt 1621 aagtttcgtg gtcatttttt gattgaggga gatttaaatc ctgataatag tgatgtggac 1681 aaactattta tccagttggt acaaacctac aatcaattat ttgaagaaaa ccctattaac 1741 gcaagtggag tagatgctaa agcgattctt tctgcacgat tgagtaaatc aagacgatta 1801 gaaaatctca ttgctcagct ccccggtgag aagaaaaatg gcttatttgg gaatctcatt 1861 gctttgtcat tgggtttgac ccctaatttt aaatcaaatt ttgatttggc agaagatgct 1921 aaattacagc tttcaaaaga tacttacgat gatgatttag ataatttatt ggcgcaaatt 1981 ggagatcaat atgctgattt gtttttggca gctaagaatt tatcagatgc tattttactt 2041 tcagatatcc taagagtaaa tactgaaata actaaggctc ccctatcagc ttcaatgatt 2101 aaacgctacg atgaacatca tcaagacttg actcttttaa aagctttagt tcgacaacaa 2161 cttccagaaa agtataaaga aatctttttt gatcaatcaa aaaacggata tgcaggttat 2221 attgatgggg gagctagcca agaagaattt tataaattta tcaaaccaat tttagaaaaa 2281 atggatggta ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa 2341 cggacctttg acaacggctc tattccccat caaattcact tgggtgagct gcatgctatt 2401 ttgagaagac aagaagactt ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa 2461 atcttgactt ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt 2521 gcatggatga ctcggaagtc tgaagaaaca attaccccat ggaattttga agaagttgtc 2581 gataaaggtg cttcagctca atcatttatt gaacgcatga caaactttga taaaaatctt 2641 ccaaatgaaa aagtactacc aaaacatagt ttgctttatg agtattttac ggtttataac 2701 gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt 2761 gaacagaaga aagccattgt tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag 2821 caattaaaag aagattattt caaaaaaata gaatgttttg atagtgttga aatttcagga 2881 gttgaagata gatttaatgc ttcattaggt acctaccatg atttgctaaa aattattaaa 2941 gataaagatt ttttggataa tgaagaaaat gaagatatct tagaggatat tgttttaaca 3001 ttgaccttat ttgaagatag ggagatgatt gaggaaagac ttaaaacata tgctcacctc 3061 tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt atactggttg gggacgtttg 3121 tctcgaaaat tgattaatgg tattagggat aagcaatctg gcaaaacaat attagatttt 3181 ttgaaatcag atggttttgc caatcgcaat tttatgcagc tgatccatga tgatagtttg 3241 acatttaaag aagacattca aaaagcacaa gtgtctggac aaggcgatag tttacatgaa 3301 catattgcaa atttagctgg tagccctgct attaaaaaag gtattttaca gactgtaaaa 3361 gttgttgatg aattggtcaa agtaatgggg cggcataagc cagaaaatat cgttattgaa 3421 atggcacgtg aaaatcagac aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa 3481 cgaatcgaag aaggtatcaa agaattagga agtcagattc ttaaagagca tcctgttgaa 3541 aatactcaat tgcaaaatga aaagctctat ctctattatc tccaaaatgg aagagacatg 3601 tatgtggacc aagaattaga tattaatcgt ttaagtgatt atgatgtcga tcacattgtt 3661 ccacaaagtt tccttaaaga cgattcaata gacaataagg tcttaacgcg ttctgataaa 3721 aatcgtggta aatcggataa cgttccaagt gaagaagtag tcaaaaagat gaaaaactat 3781 tggagacaac ttctaaacgc caagttaatc actcaacgta agtttgataa tttaacgaaa 3841 gctgaacgtg gaggtttgag tgaacttgat aaagctggtt ttatcaaacg ccaattggtt 3901 gaaactcgcc aaatcactaa gcatgtggca caaattttgg atagtcgcat gaatactaaa 3961 tacgatgaaa atgataaact tattcgagag gttaaagtga ttaccttaaa atctaaatta 4021 gtttctgact tccgaaaaga tttccaattc tataaagtac gtgagattaa caattaccat 4081 catgcccatg atgcgtatct aaatgccgtc gttggaactg ctttgattaa gaaatatcca 4141 aaacttgaat cggagtttgt ctatggtgat tataaagttt atgatgttcg taaaatgatt 4201 gctaagtctg agcaagaaat aggcaaagca accgcaaaat atttctttta ctctaatatc 4261 atgaacttct tcaaaacaga aattacactt gcaaatggag agattcgcaa acgccctcta 4321 atcgaaacta atggggaaac tggagaaatt gtctgggata aagggcgaga ttttgccaca 4381 gtgcgcaaag tattgtccat gccccaagtc aatattgtca agaaaacaga agtacagaca 4441 ggcggattct ccaaggagtc aattttacca aaaagaaatt cggacaagct tattgctcgt 4501 aaaaaagact gggatccaaa aaaatatggt ggttttgata gtccaacggt agcttattca 4561 gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag 4621 ttactaggga tcacaattat ggaaagaagt tcctttgaaa aaaatccgat tgacttttta 4681 gaagctaaag gatataagga agttaaaaaa gacttaatca ttaaactacc taaatatagt 4741 ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga attacaaaaa 4801 ggaaatgagc tggctctgcc aagcaaatat gtgaattttt tatatttagc tagtcattat 4861 gaaaagttga agggtagtcc agaagataac gaacaaaaac aattgtttgt ggagcagcat 4921 aagcattatt tagatgagat tattgagcaa atcagtgaat tttctaagcg tgttatttta 4981 gcagatgcca atttagataa agttcttagt gcatataaca aacatagaga caaaccaata 5041 cgtgaacaag cagaaaatat tattcattta tttacgttga cgaatcttgg agctcccgct 5101 gcttttaaat attttgatac aacaattgat cgtaaacgat atacgtctac aaaagaagtt 5161 ttagatgcca ctcttatcca tcaatccatc actggtcttt atgaaacacg cattgatttg 5221 agtcagctag gaggtgacgg gtcacctaag aaaaaacgaa aagttgagga tcctaaaaag 5281 aaacgaaaag ttgataccgg ttctaaaggt gaagaactgt tcaccggtgt tgttccgatc 5341 ctggttgaac tggatggtga tgttaacggc cacaaattct ctgttcgtgg tgaaggtgaa 5401 ggtgatgcaa ccaacggtaa actgaccctg aaattcatct gcactaccgg taaactgccg 5461 gttccatggc cgactctggt gactaccctg acctatggtg ttcagtgttt ttctcgttac 5521 ccggatcaca tgaagcagca tgatttcttc aaatctgcaa tgccggaagg ttatgtacag 5581 gagcgcacca tttctttcaa agacgatggc acctacaaaa cccgtgcaga ggttaaattt 5641 gaaggtgata ctctggtgaa ccgtattgaa ctgaaaggca ttgatttcaa agaggacggc 5701 aacatcctgg gccacaaact ggaatataac ttcaactccc ataacgttta catcaccgca 5761 gacaaacaga agaacggtat caaagctaac ttcaaaattc gccataacgt tgaagacggt 5821 agcgtacagc tggcggacca ctaccagcag aacactccga tcggtgatgg tccggttctg 5881 ctgccggata accactacct gtccacccag tctgttctgt ccaaagaccc gaacgaaaag 5941 cgcgaccaca tggtgctgct ggagttcgtt actgcagcag gtatcacgca cggcatggat 6001 gagctctaca aataataaca ttggaagtgg ataacggatc cgcgatcgcg gcgcgccacc 6061 tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga 6121 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa 6181 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc cacaggacgg 6241 gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg 6301 ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca 6361 acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa tggacgatat 6421 cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga 6481 cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt 6541 agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat 6601 gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 6661 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 6721 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 6781 taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 6841 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 6901 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 6961 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 7021 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc 7081 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 7141 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 7201 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 7261 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 7321 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc 7381 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 7441 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 7501 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca 7561 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 7621 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 7681 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 7741 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 7801 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 7861 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 7921 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 7981 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 8041 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 8101 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 8161 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 8221 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 8281 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 8341 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 8401 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 8461 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 8521 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc 8581 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt 8641 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg 8701 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc 8761 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 8821 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 8881 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca 8941 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg 9001 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc 9061 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct 9121 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca 9181 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt 9241 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg 9301 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc 9361 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt 9421 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg 9481 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga 9541 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt 9601 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa 9661 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg 9721 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg 9781 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa 9841 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc 9901 tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc 9961 ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg 10021 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca 10081 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc 10141 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc 10201 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc 10261 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgacgctct 10321 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc 10381 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg 10441 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct 10501 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg 10561 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac 10621 tcactatagg gagaccacaa cggtttccct ctagtgccgg ctccggagag ctctttaatt 10681 aagcggccgc cctgcaggac tcgagttcta gaaataattt tgtttaactt taagaaggag 10741 atatacatat gaaatcttct caccatcacc atcaccatgg ttcttctatg aaaatcgaag 10801 aaggtaaact ggtaatctgg attaacggcg ataaaggcta taac //