LOCUS Exported 10976 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For expression of modified SpyCas9 in e.coli.. ACCESSION . VERSION . KEYWORDS 4xNLS-pMJ915v2-sfGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10976) 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 10976) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10976 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 3718..3738 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3745..3765 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(4614..4632) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4628..4675 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(5041..5069) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(5096..5114) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5182..5286 /gene="bla" /label=AmpR promoter CDS 5287..6147 /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(5505..5524) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6318..6906 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6807..6826 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7060..7077 /label=L4440 /note="L4440 vector, forward primer" misc_feature 7092..7231 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(7157..7176) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7317..7339 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(7333..7524) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 8982..9001 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 9085..9104 /label=T7 /note="T7 promoter, forward primer" promoter 9085..9103 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 9192..9214 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 9234..9251 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 9261..10361 /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 10335..10358 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 10416..10436 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 10446..10466 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10476..10496 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10506..10526 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10536..10556 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10584..3711 /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" ORIGIN 1 tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa agcggatttg 61 cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt tttgattgag 121 ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt ggtacaaacc 181 tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc taaagcgatt 241 ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca gctccccggt 301 gagaagaaaa atggcttatt tgggaatctc attgctttgt cattgggttt gacccctaat 361 tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa agatacttac 421 gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga tttgtttttg 481 gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt aaatactgaa 541 ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca tcatcaagac 601 ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa agaaatcttt 661 tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag ccaagaagaa 721 ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga attattggtg 781 aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg ctctattccc 841 catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga cttttatcca 901 tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat tccttattat 961 gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa gtctgaagaa 1021 acaattaccc catggaattt tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt 1081 attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact accaaaacat 1141 agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa atatgttact 1201 gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat tgttgattta 1261 ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa 1321 atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa tgcttcatta 1381 ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga taatgaagaa 1441 aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga tagggagatg 1501 attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat gaaacagctt 1561 aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa tggtattagg 1621 gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt tgccaatcgc 1681 aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat tcaaaaagca 1741 caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc tggtagccct 1801 gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt caaagtaatg 1861 gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca gacaactcaa 1921 aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat caaagaatta 1981 ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc 2041 tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt agatattaat 2101 cgtttaagtg attatgatgt cgatcacatt gttccacaaa gtttccttaa agacgattca 2161 atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca 2221 agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa cgccaagtta 2281 atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt gagtgaactt 2341 gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac taagcatgtg 2401 gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa acttattcga 2461 gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa agatttccaa 2521 ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta tctaaatgcc 2581 gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt tgtctatggt 2641 gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa 2701 gcaaccgcaa aatatttctt ttactctaat atcatgaact tcttcaaaac agaaattaca 2761 cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga aactggagaa 2821 attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc catgccccaa 2881 gtcaatattg tcaagaaaac agaagtacag acaggcggat tctccaagga gtcaatttta 2941 ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc aaaaaaatat 3001 ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa 3061 gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat tatggaaaga 3121 agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa ggaagttaaa 3181 aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa cggtcgtaaa 3241 cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct gccaagcaaa 3301 tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag tccagaagat 3361 aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga gattattgag 3421 caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga taaagttctt 3481 agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa tattattcat 3541 ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga tacaacaatt 3601 gatcgtaaac gatatacgtc tacaaaagaa gttttagatg ccactcttat ccatcaatcc 3661 atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga cgggtcacct 3721 aagaaaaaac gaaaagttga ggatcctaaa aagaaacgaa aagttgatac cggttctaaa 3781 ggtgaagaac tgttcaccgg tgttgttccg atcctggttg aactggatgg tgatgttaac 3841 ggccacaaat tctctgttcg tggtgaaggt gaaggtgatg caaccaacgg taaactgacc 3901 ctgaaattca tctgcactac cggtaaactg ccggttccat ggccgactct ggtgactacc 3961 ctgacctatg gtgttcagtg tttttctcgt tacccggatc acatgaagca gcatgatttc 4021 ttcaaatctg caatgccgga aggttatgta caggagcgca ccatttcttt caaagacgat 4081 ggcacctaca aaacccgtgc agaggttaaa tttgaaggtg atactctggt gaaccgtatt 4141 gaactgaaag gcattgattt caaagaggac ggcaacatcc tgggccacaa actggaatat 4201 aacttcaact cccataacgt ttacatcacc gcagacaaac agaagaacgg tatcaaagct 4261 aacttcaaaa ttcgccataa cgttgaagac ggtagcgtac agctggcgga ccactaccag 4321 cagaacactc cgatcggtga tggtccggtt ctgctgccgg ataaccacta cctgtccacc 4381 cagtctgttc tgtccaaaga cccgaacgaa aagcgcgacc acatggtgct gctggagttc 4441 gttactgcag caggtatcac gcacggcatg gatgagctct acaaataata acattggaag 4501 tggataacgg atccgcgatc gcggcgcgcc acctggtggc cggccggtac cacgcgtgcg 4561 cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga 4621 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa 4681 aggaggaact atatccggat atccacagga cgggtgtggt cgccatgatc gcgtagtcga 4741 tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg 4801 ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat 4861 agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca 4921 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat 4981 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc 5041 attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc 5101 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 5161 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 5221 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataaca ttgaaaaagg 5281 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 5341 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 5401 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 5461 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 5521 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 5581 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 5641 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 5701 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 5761 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 5821 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 5881 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 5941 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 6001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 6061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 6121 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 6181 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 6241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 6301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 6361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 6421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 6481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 6541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 6601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 6661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 6721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 6781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 6841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 6901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 6961 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 7021 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 7081 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 7141 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc 7201 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc 7261 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 7321 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta 7381 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag 7441 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag 7501 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg 7561 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca 7621 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca 7681 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca 7741 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt 7801 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt 7861 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg 7921 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat 7981 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat 8041 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa 8101 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc 8161 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg 8221 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa 8281 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat 8341 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg 8401 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc 8461 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc 8521 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc 8581 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc 8641 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag 8701 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg 8761 ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg cattaggaag 8821 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag 8881 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa 8941 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag 9001 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg 9061 atcgagatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc 9121 cctctagtgc cggctccgga gagctcttta attaagcggc cgccctgcag gactcgagtt 9181 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgaaatct tctcaccatc 9241 accatcacca tggttcttct atgaaaatcg aagaaggtaa actggtaatc tggattaacg 9301 gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa 9361 ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa 9421 ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat 9481 ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta 9541 cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt 9601 tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc 9661 cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag 9721 aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa 9781 acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga 9841 ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg 9901 cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt 9961 ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc 10021 aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca 10081 aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg 10141 ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga 10201 aagatccacg tattgccgcc actatggaaa acgcccagaa aggtgaaatc atgccgaaca 10261 tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg 10321 gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taattcgagc tcgaacaaca 10381 acaacaataa caataacaac aacctcggga tcgaggaaaa cctgtacttc caatccaatg 10441 ccacccccaa gaagaagcga aaggtgggtg ggtccccaaa gaagaaacgg aaagtaggcg 10501 gctcccccaa aaagaagcga aaagtagggg gtagccccaa gaagaagcgg aaggtaggta 10561 tccacggagt cccagcagct accatggata agaaatactc aataggctta gatatcggca 10621 caaatagcgt cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca 10681 aggttctggg aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat 10741 ttgacagtgg agagacagcg gaagcgactc gtctcaaacg gacagctcgt agaaggtata 10801 cacgtcggaa gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag 10861 tagatgatag tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc 10921 atgaacgtca tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaa //