LOCUS Exported 11752 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Plasmid for constituive spCas9 and tet-inducible GAL7 targeting sgRNA expression for double stranded break formation in yeast. ACCESSION . VERSION . KEYWORDS pNSU299_GAL7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11752) TITLE Burgess Lab Plasmids REFERENCE 2 (bases 1 to 11752) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11752 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 3114..3134 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 3150..3397 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(3166..3184) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(3405..3424) /label=T7 /note="T7 promoter, forward primer" promoter complement(3406..3424) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3434..3451) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3434..3450) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3443..3465) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3595..4050 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3682..3701) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3892..3913 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(4147..4821) /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(4822..5102) /gene="S. cerevisiae TRP1" /label=TRP1 promoter primer_bind complement(5117..5136) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5236..5258 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5296..5314) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5382..5486 /gene="bla" /label=AmpR promoter CDS 5487..6347 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5705..5724) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6518..7106 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7007..7026 /label=pBR322ori-F /note="pBR322 origin, forward primer" protein_bind 7490..7508 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" misc_RNA 7594..7669 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 7682..7744 /label=RPR1 terminator /note="transcription terminator for the S. cerevisiae RNase P RNA gene" CDS 8895..9518 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind complement(8968..8987) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind 10015..10032 /label=L4440 /note="L4440 vector, forward primer" promoter 10185..10215 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10223..10239 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 10228..10250 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 10247..10263 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 10247..10263 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 10282..10302 /label=T3 /note="T3 promoter, forward primer" promoter 10284..10302 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 10320..10718 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind 10722..10738 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 10722..10738 /label=pBluescriptSK /note="For pBluescript vector" CDS 10750..3101 /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 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt 61 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat 121 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca 181 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc 241 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag 301 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc 361 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc 421 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga 481 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt 541 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga 601 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga 661 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt 721 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc 781 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca 841 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac 901 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc 961 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga 1021 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc 1081 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg 1141 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa 1201 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata 1261 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga 1321 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa 1381 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact 1441 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg 1501 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata 1561 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag 1621 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac 1681 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg 1741 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc 1801 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag 1861 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg 1921 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca 1981 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag 2041 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta 2101 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg 2161 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg 2221 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg 2281 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga 2341 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg 2401 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta 2461 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg 2521 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca 2581 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg 2641 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt 2701 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga 2761 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg 2821 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca 2881 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc 2941 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc 3001 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc 3061 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga 3121 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc 3181 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta 3241 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat 3301 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct 3361 tgagaaggtt ttgggacgct cgaaggcttt aatttgccaa ttcgccctat agtgagtcgt 3421 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta 3481 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg 3541 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc 3601 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3661 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3721 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 3781 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3841 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3901 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 3961 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 4021 attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc ttacgcatct 4081 gtgcggtatt tcacaccgca ggcaagtgca caaacaatac ttaaataaat actactcagt 4141 aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt cttttacacc 4201 atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc taagcgcatc 4261 accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagcttt cggggctctc 4321 ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg tcccacctgc 4381 ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac tgagtagtat 4441 gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga actcttggta 4501 ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat cattgaccag 4561 agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt tcggagtgcc 4621 tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa ccgggtcaat 4681 tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg aatctagagc 4741 acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac cagaactacc 4801 tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca cgtatactca 4861 cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt tttcgaccga 4921 attaattctt aatcggcaaa aaaagaaaag ctccggatca agattgtacg taaggtgaca 4981 agctattttt caataaagaa tatcttccac tactgccatc tggcgtcata actgcaaagt 5041 acacatatat tacgatgctg tctattaaat gcttcctata ttatatatat agtaatgtcg 5101 tttggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc 5161 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac 5221 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac 5281 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa 5341 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 5401 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 5461 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 5521 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 5581 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 5641 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 5701 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 5761 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 5821 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 5881 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct ttttttcaca 5941 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 6001 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 6061 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 6121 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 6181 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 6241 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 6301 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 6361 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 6421 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 6481 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 6541 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 6601 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 6661 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 6721 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 6781 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 6841 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 6901 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 6961 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 7021 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 7081 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 7141 ccttttgctg gccttttgct cgggggatct gccaattgaa cataacatgg tagttacata 7201 tactagtaat atggttcggc acacattaaa agtataaaaa ctatctgaat tacgaattac 7261 atatattggt cataaaaatc aatcaatcat cgtgtgtttt atatgtctct tatctaagta 7321 taagaatatc catagttaat attcacttac gctacctttt aacctgtaat cattgtcaac 7381 aggatatgtt aacgacccac attgataaac gctagtattt ctttttcctc ttcttattgg 7441 ccggctgtct ctatactccc ctatagtctg tttcttttcg tttcgattgt ccctatcagt 7501 gatagagatg gcgcacatgg tacgctgtgg tgctcgcggc tgggaacgaa actctgggag 7561 ctgcgattgg caggcctcct gttgacctaa ccagttttag agctagaaat agcaagttaa 7621 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttctcga 7681 gccatatcca acttccaatt taatctttct tttttaattt tcacttattt gcgatacaga 7741 aagaaaaaag cgatagtaac tattgaattt tgtttggatt tggttagatt agatatggtt 7801 tctctttata tttacatgct aaaaatgggc tacaccagag atacataatt agatatatat 7861 acgccagtac accttatcgg cccaagcctt gtcccaaggc agcgttttgt tcttggaaac 7921 gctgccctac acgttcgcta tgcttcaaga acttttctga gcacttcatg atgcatgttt 7981 gttccttatt ggttagcttt gatgttgtga agtcattgac acagtctgtg aaacatcttt 8041 ctaccagatt agagtacaaa cgcatgaaat ccttcatttg cttttgttcc actacttttt 8101 ggaactcttg ttgttctttc ctgcaggtag tcgtgcaatg tatgacttta agatttgtga 8161 gcaggaagaa aagggagaat cttctaacga taaacccttg aaaaactggg tagactacgc 8221 tatgttgagt tgctacgcag gctgcacaat tacacgagaa tgctcccgcc taggatttaa 8281 ggctaaggga cgtgcaatgc agacgacaga tctaaatgac cgtgtcggtg aagtgttcgc 8341 caaacttttc ggttaacaca tgcagtgatg cacgcgcgat ggtgctaagt tacatatata 8401 tatatatata tatatatata tatatatagc catagtgatg tctaagtaac ctttatggta 8461 tatttcttaa tgtggaaaga tactagcgcg cgcacccaca cacaagcttc gtcttttctt 8521 gaagaaaaga ggaagctcgc taaatgggat tccactttcc gttccctgcc agctgatgga 8581 aaaaggttag tggaacgatg aagaataaaa agagagatcc actgaggtga aatttcagct 8641 gacagcgagt ttcatgatcg tgatgaacaa tggtaacgag ttgtggctgt tgccagggag 8701 ggtggttctc aacttttaat gtatggccaa atcgctactt gggtttgtta tataacaaag 8761 aagaaataat gaactgattc tcttcctcct tcttgtcctt tcttaattct gttgtaatta 8821 ccttcctttg taattttttt tgtaattatt cttcttaata atccaaacaa acacacatat 8881 tacaatatat taaaatgtct agattagata aaagtaaagt gattaacagc gcattagagc 8941 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg 9001 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag 9061 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc 9121 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg 9181 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc 9241 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg 9301 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag 9361 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat 9421 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg 9481 gattagaaaa acaacttaaa tgtgaaagtg ggtcttaagt ctgaagaatg aatgatttga 9541 tgatttcttt ttccctccat ttttcttact gaatatatca atgatataga cttgtatagt 9601 ttattatttc aaattaagta gctatatata gtcaagataa cgtttgtttg acacgattac 9661 attattcgtc gacatctttt ttcagcctgt cgtggtagca atttgaggag tattattaat 9721 tgaataggtt cattttgcgc tcgcataaac agttttcgtc agggacagta tgttggaatg 9781 agtggtaatt aatggtgaca tgacatgtta tagcaataac cttgatgttt acatcgtagt 9841 ttaatgtaca ccccgcgaat tcgttcaagt aggagtgcac caattgcaaa gggaaaagct 9901 gaatgggcag ttcgaaccgc ggtctttcct gcgttatccc ctgattctgt ggataaccgt 9961 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag 10021 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg 10081 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 10141 aacgcaatta atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt 10201 ccggctccta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat 10261 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc 10321 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca 10381 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc 10441 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt 10501 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat 10561 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg 10621 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc 10681 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttctagaact agtggatccc 10741 ccgggaaaaa tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggt 10801 tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat 10861 accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag 10921 acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat 10981 cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc 11041 ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca 11101 atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat 11161 ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg 11221 ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac 11281 aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa 11341 gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc 11401 aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg 11461 tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac 11521 ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat 11581 ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca 11641 gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg 11701 agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt gc //