LOCUS Exported 10133 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION For expression of modified SpyCas9 in e.coli.. ACCESSION . VERSION . KEYWORDS pMJ915v2+Nterm Spe1 +Cterm Age1 site SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10133) 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 10133) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10133 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 97..116 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 350..367 /label=L4440 /note="L4440 vector, forward primer" misc_feature 382..521 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(447..466) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 607..629 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(623..814) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 2272..2291 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 2375..2394 /label=T7 /note="T7 promoter, forward primer" promoter 2375..2393 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 2482..2504 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2524..2541 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2551..3651 /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 3625..3648 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 3706..3726 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 3742..7845 /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 7852..7872 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7879..7899 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(8037..8055) /label=T7 Term /note="T7 terminator, reverse primer" terminator 8051..8098 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(8464..8492) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(8519..8537) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8605..8709 /gene="bla" /label=AmpR promoter CDS 8710..9570 /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(8928..8947) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9741..196 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 61 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 361 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 421 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 481 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac 541 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 601 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc 661 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat 721 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg 781 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc 841 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg 901 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc 961 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag 1021 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg 1081 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg 1141 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg 1201 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga 1261 gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc 1321 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag 1381 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc 1441 gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag 1501 gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga 1561 ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc 1621 cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc 1681 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat 1741 ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc 1801 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc 1861 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg 1921 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat 1981 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga 2041 gctgactggg ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg 2101 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg 2161 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac 2221 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc 2281 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc 2341 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat agggagacca 2401 caacggtttc cctctagtgc cggctccgga gagctcttta attaagcggc cgccctgcag 2461 gactcgagtt ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgaaatct 2521 tctcaccatc accatcacca tggttcttct atgaaaatcg aagaaggtaa actggtaatc 2581 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa 2641 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag 2701 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc 2761 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg 2821 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct 2881 gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg 2941 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc 3001 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc 3061 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa 3121 gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat 3181 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg 3241 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc 3301 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc 3361 agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt 3421 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa 3481 gagttggcga aagatccacg tattgccgcc actatggaaa acgcccagaa aggtgaaatc 3541 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac 3601 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taattcgagc 3661 tcgaacaaca acaacaataa caataacaac aacctcggga tcgaggaaaa cctgtacttc 3721 caatccaatg ccactagtac catggataag aaatactcaa taggcttaga tatcggcaca 3781 aatagcgtcg gatgggcggt gatcactgat gaatataagg ttccgtctaa aaagttcaag 3841 gttctgggaa atacagaccg ccacagtatc aaaaaaaatc ttataggggc tcttttattt 3901 gacagtggag agacagcgga agcgactcgt ctcaaacgga cagctcgtag aaggtataca 3961 cgtcggaaga atcgtatttg ttatctacag gagatttttt caaatgagat ggcgaaagta 4021 gatgatagtt tctttcatcg acttgaagag tcttttttgg tggaagaaga caagaagcat 4081 gaacgtcatc ctatttttgg aaatatagta gatgaagttg cttatcatga gaaatatcca 4141 actatctatc atctgcgaaa aaaattggta gattctactg ataaagcgga tttgcgctta 4201 atctatttgg ccttagcgca tatgattaag tttcgtggtc attttttgat tgagggagat 4261 ttaaatcctg ataatagtga tgtggacaaa ctatttatcc agttggtaca aacctacaat 4321 caattatttg aagaaaaccc tattaacgca agtggagtag atgctaaagc gattctttct 4381 gcacgattga gtaaatcaag acgattagaa aatctcattg ctcagctccc cggtgagaag 4441 aaaaatggct tatttgggaa tctcattgct ttgtcattgg gtttgacccc taattttaaa 4501 tcaaattttg atttggcaga agatgctaaa ttacagcttt caaaagatac ttacgatgat 4561 gatttagata atttattggc gcaaattgga gatcaatatg ctgatttgtt tttggcagct 4621 aagaatttat cagatgctat tttactttca gatatcctaa gagtaaatac tgaaataact 4681 aaggctcccc tatcagcttc aatgattaaa cgctacgatg aacatcatca agacttgact 4741 cttttaaaag ctttagttcg acaacaactt ccagaaaagt ataaagaaat cttttttgat 4801 caatcaaaaa acggatatgc aggttatatt gatgggggag ctagccaaga agaattttat 4861 aaatttatca aaccaatttt agaaaaaatg gatggtactg aggaattatt ggtgaaacta 4921 aatcgtgaag atttgctgcg caagcaacgg acctttgaca acggctctat tccccatcaa 4981 attcacttgg gtgagctgca tgctattttg agaagacaag aagactttta tccattttta 5041 aaagacaatc gtgagaagat tgaaaaaatc ttgacttttc gaattcctta ttatgttggt 5101 ccattggcgc gtggcaatag tcgttttgca tggatgactc ggaagtctga agaaacaatt 5161 accccatgga attttgaaga agttgtcgat aaaggtgctt cagctcaatc atttattgaa 5221 cgcatgacaa actttgataa aaatcttcca aatgaaaaag tactaccaaa acatagtttg 5281 ctttatgagt attttacggt ttataacgaa ttgacaaagg tcaaatatgt tactgaagga 5341 atgcgaaaac cagcatttct ttcaggtgaa cagaagaaag ccattgttga tttactcttc 5401 aaaacaaatc gaaaagtaac cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa 5461 tgttttgata gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc attaggtacc 5521 taccatgatt tgctaaaaat tattaaagat aaagattttt tggataatga agaaaatgaa 5581 gatatcttag aggatattgt tttaacattg accttatttg aagataggga gatgattgag 5641 gaaagactta aaacatatgc tcacctcttt gatgataagg tgatgaaaca gcttaaacgt 5701 cgccgttata ctggttgggg acgtttgtct cgaaaattga ttaatggtat tagggataag 5761 caatctggca aaacaatatt agattttttg aaatcagatg gttttgccaa tcgcaatttt 5821 atgcagctga tccatgatga tagtttgaca tttaaagaag acattcaaaa agcacaagtg 5881 tctggacaag gcgatagttt acatgaacat attgcaaatt tagctggtag ccctgctatt 5941 aaaaaaggta ttttacagac tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg 6001 cataagccag aaaatatcgt tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc 6061 cagaaaaatt cgcgagagcg tatgaaacga atcgaagaag gtatcaaaga attaggaagt 6121 cagattctta aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa gctctatctc 6181 tattatctcc aaaatggaag agacatgtat gtggaccaag aattagatat taatcgttta 6241 agtgattatg atgtcgatca cattgttcca caaagtttcc ttaaagacga ttcaatagac 6301 aataaggtct taacgcgttc tgataaaaat cgtggtaaat cggataacgt tccaagtgaa 6361 gaagtagtca aaaagatgaa aaactattgg agacaacttc taaacgccaa gttaatcact 6421 caacgtaagt ttgataattt aacgaaagct gaacgtggag gtttgagtga acttgataaa 6481 gctggtttta tcaaacgcca attggttgaa actcgccaaa tcactaagca tgtggcacaa 6541 attttggata gtcgcatgaa tactaaatac gatgaaaatg ataaacttat tcgagaggtt 6601 aaagtgatta ccttaaaatc taaattagtt tctgacttcc gaaaagattt ccaattctat 6661 aaagtacgtg agattaacaa ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt 6721 ggaactgctt tgattaagaa atatccaaaa cttgaatcgg agtttgtcta tggtgattat 6781 aaagtttatg atgttcgtaa aatgattgct aagtctgagc aagaaatagg caaagcaacc 6841 gcaaaatatt tcttttactc taatatcatg aacttcttca aaacagaaat tacacttgca 6901 aatggagaga ttcgcaaacg ccctctaatc gaaactaatg gggaaactgg agaaattgtc 6961 tgggataaag ggcgagattt tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat 7021 attgtcaaga aaacagaagt acagacaggc ggattctcca aggagtcaat tttaccaaaa 7081 agaaattcgg acaagcttat tgctcgtaaa aaagactggg atccaaaaaa atatggtggt 7141 tttgatagtc caacggtagc ttattcagtc ctagtggttg ctaaggtgga aaaagggaaa 7201 tcgaagaagt taaaatccgt taaagagtta ctagggatca caattatgga aagaagttcc 7261 tttgaaaaaa atccgattga ctttttagaa gctaaaggat ataaggaagt taaaaaagac 7321 ttaatcatta aactacctaa atatagtctt tttgagttag aaaacggtcg taaacggatg 7381 ctggctagtg ccggagaatt acaaaaagga aatgagctgg ctctgccaag caaatatgtg 7441 aattttttat atttagctag tcattatgaa aagttgaagg gtagtccaga agataacgaa 7501 caaaaacaat tgtttgtgga gcagcataag cattatttag atgagattat tgagcaaatc 7561 agtgaatttt ctaagcgtgt tattttagca gatgccaatt tagataaagt tcttagtgca 7621 tataacaaac atagagacaa accaatacgt gaacaagcag aaaatattat tcatttattt 7681 acgttgacga atcttggagc tcccgctgct tttaaatatt ttgatacaac aattgatcgt 7741 aaacgatata cgtctacaaa agaagtttta gatgccactc ttatccatca atccatcact 7801 ggtctttatg aaacacgcat tgatttgagt cagctaggag gtgacgggtc acctaagaaa 7861 aaacgaaaag ttgaggatcc taaaaagaaa cgaaaagttg ataccggtta ataacattgg 7921 aagtggataa cggatccgcg atcgcggcgc gccacctggt ggccggccgg taccacgcgt 7981 gcgcgctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc 8041 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct 8101 gaaaggagga actatatccg gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt 8161 cgatagtggc tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca 8221 gtgctccgag aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc 8281 catagtgact ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg 8341 gcataaccaa gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg 8401 cattgttaga tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac 8461 cgcattaaag cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc 8521 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca 8581 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 8641 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata acattgaaaa 8701 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt 8761 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag 8821 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt 8881 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg 8941 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag 9001 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta 9061 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg 9121 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta 9181 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac 9241 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt 9301 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca 9361 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag 9421 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta 9481 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag 9541 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt 9601 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 9661 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 9721 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 9781 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 9841 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 9901 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 9961 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 10021 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 10081 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gct //