LOCUS Exported 11928 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION High copy Cas9 expression vector with LEU2 marker for yeast transformation. ACCESSION . VERSION . KEYWORDS pJH001 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11928) AUTHORS Laughery MF, Hunter T, Brown A, Hoopes J, Ostbye T, Shumaker T, Wyrick JJ TITLE New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. JOURNAL Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. PUBMED 26305040 REFERENCE 2 (bases 1 to 11928) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11928 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS complement(664..1758) /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" promoter complement(1759..2166) /gene="S. cerevisiae LEU2" /label=LEU2 promoter rep_origin complement(2467..2922) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2604..2625) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2816..2835 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3048..3070 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3062..3079 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3063..3079 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3089..3108 /label=T7 /note="T7 promoter, forward primer" promoter 3089..3107 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 3132..3148 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 3133..3149 /label=KS primer /note="common sequencing primer, one of multiple similar variants" promoter 3194..3860 /gene="S. cerevisiae TDH3" /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase" primer_bind 3730..3753 /label=GPDpro-F /note="S. cerevisiae GPD promoter, forward primer" regulatory 3889..3898 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3895..7998 /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 8008..8028 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 8056..8243 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(8111..8135) /label=pHybLex-R /note="ADH terminator, reverse primer" primer_bind complement(8323..8343) /label=T3 /note="T3 promoter, forward primer" promoter complement(8323..8341) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(8362..8378) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8362..8378) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8375..8397) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8386..8402 /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)." promoter complement(8410..8440) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8455..8476 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8593..8610) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8764..9352) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8844..8863) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9523..10383) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 10146..10165 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10384..10488) /gene="bla" /label=AmpR promoter rep_origin 10515..11861 /label=2u ori /note="yeast 2u plasmid origin of replication" primer_bind 11901..11919 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt 241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc 301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa 361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct 421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta 481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg 541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc 601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc 661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact 721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc 781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga 841 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca 901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg 961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat 1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc 1081 agaatcaatc aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc 1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat 1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac 1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct 1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc 1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct 1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac 1501 actacctgta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt 1561 ggaggcttcc agcgcctcat ctggaagtgg aacacctgta gcatctatag cagcaccacc 1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag 1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat 1741 cttcttaggg gcagacatta gaatggtata tccttgaaat atatatatat atattgctga 1801 aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa 1861 caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg 1921 aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc 1981 caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt 2041 ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga 2101 aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc 2161 acagttaact gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca 2221 accaattact tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac 2281 acatgaacaa ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt 2341 gttgggattc catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct 2401 cctcgaccgg tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 2461 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct 2941 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 3001 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 3061 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac 3121 cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattca gtctactagt 3181 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt 3241 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt 3301 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga 3361 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag 3421 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt 3481 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac 3541 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc 3601 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa 3661 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt 3721 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat 3781 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata 3841 aacacacata aacaaacaaa agatctgata tggatcgaat tagatctcgc caccatggac 3901 aagaagtatt ctatcggact ggacatcggg actaatagcg tcgggtgggc cgtgatcact 3961 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc 4021 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc 4081 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt 4141 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa 4201 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc 4261 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg 4321 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc 4381 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat 4441 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat 4501 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt 4561 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt 4621 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc 4681 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc 4741 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg 4801 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt 4861 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag 4921 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac 4981 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag 5041 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag 5101 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc 5161 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa 5221 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt 5281 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg 5341 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc 5401 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac 5461 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc 5521 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag 5581 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg 5641 gtcgaagatc ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag 5701 gacaaggatt ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact 5761 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg 5821 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt 5881 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc 5941 cttaagagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg 6001 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag 6061 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa 6121 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag 6181 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa 6241 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa 6301 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg 6361 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga ccacatcgtc 6421 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag 6481 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat 6541 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag 6601 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg 6661 gagactcggc agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag 6721 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg 6781 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat 6841 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca 6901 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt 6961 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc 7021 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc 7081 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc 7141 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc 7201 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg 7261 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct 7321 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa 7381 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc 7441 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc 7501 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag 7561 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac 7621 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat 7681 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc 7741 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt 7801 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc 7861 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg 7921 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc 7981 tcacagctcg gaggggatga gggagctccc aagaaaaagc gcaaggtagg tagttccgga 8041 tcctagactc tcgaggcgaa tttcttatga tttatgattt ttattattaa ataagttata 8101 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta 8161 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct 8221 cttattgacc acacctctac cggcatgccg agcaaatgcc tgcaaatcgc tccccatttc 8281 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat 8341 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 8401 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 8461 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 8521 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 8581 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 8641 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 8701 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 8761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 8821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 8881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 8941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 9001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 9061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 9121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 9181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 9241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 9301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 9361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 9421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 9481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 9541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 9601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 9661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 9721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 9781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 9841 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 9901 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 9961 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 10021 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 10081 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 10141 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 10201 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 10261 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 10321 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 10381 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 10441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 10501 aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga 10561 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg 10621 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa 10681 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg 10741 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa 10801 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg 10861 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa 10921 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg 10981 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta 11041 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat 11101 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat 11161 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa 11221 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag 11281 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat 11341 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt 11401 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg 11461 gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga acttcggaat 11521 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac 11581 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat 11641 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg 11701 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt 11761 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta 11821 gtctcatcct tcaatgctat catttccttt gatattggat catctaagaa accattatta 11881 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc //