LOCUS Exported 12576 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pRS416-dCas9-Mxi1+TetR+pRPR1(TetO)-NotI-gRNA plasmid marked with His, expressing dCas9-Mxi1 under Tef1 promoter, and a tet-inducible promoter for gRNA expression.. ACCESSION . VERSION . KEYWORDS ampl43 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12576) AUTHORS Momen-Roknabadi A, Oikonomou P, Zegans M, Tavazoie S TITLE An inducible CRISPR interference library for genetic interrogation of Saccharomyces cerevisiae biology. JOURNAL Commun Biol. 2020 Nov 27;3(1):723. doi: 10.1038/s42003-020-01452-9. PUBMED 33247197 REFERENCE 2 (bases 1 to 12576) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12576 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 136..154 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" misc_RNA 227..302 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 315..377 /label=RPR1 terminator /note="transcription terminator for the S. cerevisiae RNase P RNA gene" CDS 1512..2135 /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(1585..1604) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind 2632..2649 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2766..2787 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2802..2832 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2840..2856 /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 2845..2867 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2864..2880 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2864..2880 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2899..2919 /label=T3 /note="T3 promoter, forward primer" promoter 2901..2919 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2937..3335 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 3358..3378 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3388..7491 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 7513..7533 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7567..7689 /codon_start=1 /product="transcriptional repressor domain from the mouse Max-interacting protein 1 (Schreiber-Agus et al., 1995)" /label=Mxi1 /translation="MERVRMINVQRLLEAAEFLERRERECEHGYASSFPSMPSPR" terminator 7713..7960 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(7729..7747) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(7978..7997) /label=T7 /note="T7 promoter, forward primer" promoter complement(7979..7997) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8007..8024) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8007..8023) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8016..8038) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8164..8619 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8251..8270) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8461..8482 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(8818..9480) /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" promoter complement(9481..9668) /gene="S. cerevisiae HIS3" /label=HIS3 promoter primer_bind complement(9815..9834) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9934..9956 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(9994..10012) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 10060..10563 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 10593..10697 /gene="bla" /label=AmpR promoter CDS 10698..11558 /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 complement(10916..10935) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 11729..12317 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 12218..12237 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccttttaacc tgtaatcatt gtcaacagga tatgttaacg acccacattg ataaacgcta 61 gtatttcttt ttcctcttct tattggccgg ctgtctctat actcccctat agtctgtttc 121 ttttcgtttc gattgtccct atcagtgata gagatggcgc acatggtacg ctgtggtgct 181 cgcggctggg aacgaaactc tgggagctgc gattggcagc ggccgcgttt tagagctaga 241 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt 301 gcttttttct cgagccatat ccaacttcca atttaatctt tcttttttaa ttttcactta 361 tttgcgatac agaaagaaaa aagcgatagt aactattgaa ttttgtttgg atttggttag 421 attagatatg gtttctcttt atatttacat gctaaaaatg ggctacacca gagatacata 481 attagatata tatacgccag tacaccttat cggcccaagc cttgtcccaa ggcagcgttt 541 tgttcttgga aacgctgccc tacacgttcg ctatgcttca agaacttttc tgagcacttc 601 atgatgcatg tttgttcctt attggttagc tttgatgttg tgaagtcatt gacacagtct 661 gtgaaacatc tttctaccag attagagtac aaacgcatga aatccttcat ttgcttttgt 721 tccactactt tttggaactc ttgttgttct ttcctgcagg tagtcgtgca atgtatgact 781 ttaagatttg tgagcaggaa gaaaagggag aatcttctaa cgataaaccc ttgaaaaact 841 gggtagacta cgctatgttg agttgctacg caggctgcac aattacacga gaatgctccc 901 gcctaggatt taaggctaag ggacgtgcaa tgcagacgac agatctaaat gaccgtgtcg 961 gtgaagtgtt cgccaaactt ttcggttaac acatgcagtg atgcacgcgc gatggtgcta 1021 agttacatat atatatatat gtatagccat agtgatgtct aagtaacctt tatggtatat 1081 ttcttaatgt ggaaagatac tagcgcgcgc acccacacac aagcttcgtc ttttcttgaa 1141 gaaaagagga agctcgctaa atgggattcc actttccgtt ccctgccagc tgatggaaaa 1201 aggttagtgg aacgatgaag aataaaaaga gagatccact gaggtgaaat ttcagctgac 1261 agcgagtttc atgatcgtga tgaacaatgg taacgagttg tggctgttgc cagggagggt 1321 ggttctcaac ttttaatgta tggccaaatc gctacttggg tttgttatat aacaaagaag 1381 aaataatgaa ctgattctct tcctccttct tgtcctttct taattctgtt gtaattacct 1441 tcctttgtaa ttttttttgt aattattctt cttaataatc caaacaaaca cacatattac 1501 aatatattaa aatgtctaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc 1561 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag 1621 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca 1681 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag 1741 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag 1801 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat 1861 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg 1921 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag 1981 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat 2041 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat 2101 tagaaaaaca acttaaatgt gaaagtgggt cttaagtctg aagaatgaat gatttgatga 2161 tttctttttc cctccatttt tcttactgaa tatatcaatg atatagactt gtatagttta 2221 ttatttcaaa ttaagtagct atatatagtc aagataacgt ttgtttgaca cgattacatt 2281 attcgtcgac atcttttttc agcctgtcgt ggtagcaatt tgaggagtat tattaattga 2341 ataggttcat tttgcgctcg cataaacagt tttcgtcagg gacagtatgt tggaatgagt 2401 ggtaattaat ggtgacatga catgttatag caataacctt gatgtttaca tcgtagttta 2461 atgtacaccc cgcgaattcg ttcaagtagg agtgcaccaa ttgcaaaggg aaaagctgaa 2521 tgggcagttc gaaccgcggt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 2581 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 2641 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 2701 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 2761 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg 2821 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac 2881 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcata 2941 gcttcaaaat gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg 3001 ccgtaccact tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag 3061 ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct 3121 ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt 3181 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct 3241 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg 3301 ctcattagaa agaaagcata gcaatctaat ctaagttttc tagagatgtc tagagcccca 3361 aagaagaaga gaaaagttag acccgggatg gacaagaagt actccattgg gctcgctatc 3421 ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa 3481 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc 3541 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga 3601 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct 3661 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa 3721 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag 3781 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg 3841 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag 3901 ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact 3961 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc 4021 ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg 4081 gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac 4141 tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac 4201 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg 4261 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag 4321 atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac 4381 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc 4441 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa 4501 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta 4561 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc 4621 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc 4681 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat 4741 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag 4801 accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc 4861 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac 4921 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca 4981 gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc 5041 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag 5101 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg 5161 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag 5221 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg 5281 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc 5341 aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga 5401 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg 5461 aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca 5521 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca 5581 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg 5641 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag 5701 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg 5761 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc 5821 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat 5881 cggctctccg actacgacgt ggatgctatc gtgccccagt cttttctcaa agatgattct 5941 attgataata aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc 6001 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg 6061 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg 6121 gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg 6181 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga 6241 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag 6301 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca 6361 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga 6421 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag 6481 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca 6541 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa 6601 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag 6661 gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc 6721 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac 6781 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa 6841 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga 6901 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa 6961 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa 7021 cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa 7081 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat 7141 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag 7201 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt 7261 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac 7321 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata 7381 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca 7441 attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct 7501 gacgagggag ctcccaagaa aaagcgcaag gtaggtagtt ccaagcttgg cggcagcggc 7561 ggcagcatgg aacgtgtgag aatgattaat gtgcaaaggc tgttagaagc cgcagagttt 7621 ttagaaagaa gagaaagaga atgcgaacac gggtatgcca gttctttccc tagcatgccc 7681 tctcccagag gctaactcga gctcttctcg agtcatgtaa ttagttatgt cacgcttaca 7741 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt 7801 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca 7861 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt 7921 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc 7981 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa 8041 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 8101 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 8161 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 8221 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 8281 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 8341 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 8401 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 8461 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 8521 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 8581 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc ctgatgcaaa ttgaagctct 8641 aatttgtgag tttagtaaaa gaagcgcgcc tcgttcagaa tgacacgtat agaatgatgc 8701 attaccttgt catcttcagt atcatactgt tcgtatacat acttactgac attcataggt 8761 atacatatat acacatgtat atatatcgta tgctgcagct ttaaataatc ggtgtcacta 8821 cataagaaca cctttggtgg agggaacatc gttggtacca ttgggcgagg tggcttctct 8881 tatggcaacc gcaagagcct tgaacgcact ctcactacgg tgatgatcat tcttgcctcg 8941 cagacaatca acgtggaggg taattctgct agcctctgca aagctttcaa gaaaatgcgg 9001 gatcatctcg caagagagat ctcctacttt ctccctttgc aaaccaagtt cgacaactgc 9061 gtacggcctg ttcgaaagat ctaccaccgc tctggaaagt gcctcatcca aaggcgcaaa 9121 tcctgatcca aaccttttta ctccacgcac ggcccctagg gcctctttaa aagcttgacc 9181 gagagcaatc ccgcagtctt cagtggtgtg atggtcgtct atgtgtaagt caccaatgca 9241 ctcaacgatt agcgaccagc cggaatgctt ggccagagca tgtatcatat ggtccagaaa 9301 ccctatacct gtgtggacgt taatcacttg cgattgtgtg gcctgttctg ctactgcttc 9361 tgcctctttt tctgggaaga tcgagtgctc tatcgctagg ggaccaccct ttaaagagat 9421 cgcaatctga atcttggttt catttgtaat acgctttact agggctttct gctctgtcat 9481 ctttgccttc gtttatcttg cctgctcatt ttttagtata ttcttcgaag aaatcacatt 9541 actttatata atgtataatt cattatgtga taatgccaat cgctaagaaa aaaaaagagt 9601 catccgctag gtggaaaaaa aaaaatgaaa atcattaccg aggcataaaa aaatatagag 9661 tgtactagag gaggccaaga gtaatagaaa aagaaaattg cgggaaagga ctgtgttatg 9721 acttccctga ctaatgccgt gttcaaacga tacctggcag tgactcctag cgctcaccaa 9781 gctcttaaaa cgggaattta tggtgcactc tcagtacaat ctgctctgat gccgcatagt 9841 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc 9901 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt 9961 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg 10021 ttaatgtcat gataataatg gtttcttagg gtccttttca tcacgtgcta taaaaataat 10081 tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat 10141 taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca 10201 aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct 10261 tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat 10321 cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct 10381 ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta 10441 ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt 10501 aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg 10561 atccgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 10621 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 10681 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 10741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 10801 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 10861 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 10921 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 10981 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 11041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 11101 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 11161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 11221 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 11281 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 11341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 11401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 11461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 11521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 11581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 11641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 11701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 11761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 11821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 11881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 11941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 12001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 12061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 12121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 12181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 12241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 12301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 12361 ggccttttgc tggccttttg ctcgggggat ctgccaattg aacataacat ggtagttaca 12421 tatactagta atatggttcg gcacacatta aaagtataaa aactatctga attacgaatt 12481 acatatattg gtcataaaaa tcaatcaatc atcgtgtgtt ttatatgtct cttatctaag 12541 tataagaata tccatagtta atattcactt acgcta //