LOCUS Exported 14606 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses MCP-VP64 and dCas9 in Yeast cells. ACCESSION . VERSION . KEYWORDS pJZC595 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14606) AUTHORS Zalatan JG, Lee ME, Almeida R, Gilbert LA, Whitehead EH, La Russa M, Tsai JC, Weissman JS, Dueber JE, Qi LS, Lim WA TITLE Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. JOURNAL Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. PUBMED 25533786 REFERENCE 2 (bases 1 to 14606) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14606 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 276..683 /gene="S. cerevisiae LEU2" /label=LEU2 promoter promoter 3292..3996 /gene="S. cerevisiae ADH1" /label=ADH1 promoter /note="promoter for alcohol dehydrogenase 1" CDS 4024..4044 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4417..4566 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" promoter 4994..5660 /gene="S. cerevisiae TDH3" /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase" primer_bind 5530..5553 /label=GPDpro-F /note="S. cerevisiae GPD promoter, forward primer" CDS 5670..9770 /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 9783..9803 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9807..9827 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9831..9851 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(11153..11172) /label=T7 /note="T7 promoter, forward primer" promoter complement(11154..11172) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(11179..11196) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(11179..11195) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11188..11210) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 11336..11791 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11423..11442) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11633..11654 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(12147..12166) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 12266..12288 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(12326..12344) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 12429..12533 /gene="bla" /label=AmpR promoter CDS 12534..13394 /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(12752..12771) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 13565..14153 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 14054..14073 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 14307..14324 /label=L4440 /note="L4440 vector, forward primer" protein_bind 14441..14462 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 14477..14507 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 14515..14531 /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 14520..14542 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 14539..14555 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 14539..14555 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 14576..14594 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta 61 catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc aaaatccaca 121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat 181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta 241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt 301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac 361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca 421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga 481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat 541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt 601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata 661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa 721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa 781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc 841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg 901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa 961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt 1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct 1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc 1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat 1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga 1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa 1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga 1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc 1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt 1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc 1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg 1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga 1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca 1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag 1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca 1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg 1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc 1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc 2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa 2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga 2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc 2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag 2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga 2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc 2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat 2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggcccta aaacaagaag 2521 agggttgact acatcacgat gagggggatc gaagaaatga tggtaaatga aataggaaat 2581 caaggagcat gaaggcaaaa gacaaatata agggtcgaac gaaaaataaa gtgaaaagtg 2641 ttgatatgat gtatttggct ttgcggcgcc gaaaaaacga gtttacgcaa ttgcacaatc 2701 atgctgactc tgtggcggac ccgcgctctt gccggcccgg cgataacgct gggcgtgagg 2761 ctgtgcccgg cggagttttt tgcgcctgca ttttccaagg tttaccctgc gctaaggggc 2821 gagattggag aagcaataag aatgccggtt ggggttgcga tgatgacgac cacgacaact 2881 ggtgtcatta tttaagttgc cgaaagaacc tgagtgcatt tgcaacatga gtatactaga 2941 agaatgagcc aagacttgcg agacgcgagt ttgccggtgg tgcgaacaat agagcgacca 3001 tgaccttgaa ggtgagacgc gcataaccgc tagagtactt tgaagaggaa acagcaatag 3061 ggttgctacc agtataaata gacaggtaca tacaacactg gaaatggttg tctgtttgag 3121 tacgctttca attcatttgg gtgtgcactt tattatgtta caatatggaa gggaacttta 3181 cacttctcct atgcacatat attaattaaa gtccaatgct agtagagaag gggggtaaca 3241 cccctccgcg ctcttttccg atttttttct aaaccgtgga atatttcgga tatccttttg 3301 ttgtttccgg gtgtacaata tggacttcct cttttctggc aaccaaaccc atacatcggg 3361 attcctataa taccttcgtt ggtctcccta acatgtaggt ggcggagggg agatatacaa 3421 tagaacagat accagacagg acataatggg ctaaacaaga ctacaccaat tacactgcct 3481 cattgatggt ggtacataac gaactaatac tgtagcccta gacttgatag ccatcatcat 3541 atcgaagttt cactaccctt tttccatttg ccatctattg aagtaataat aggcgcatgc 3601 aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa 3661 tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca 3721 gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc 3781 ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt 3841 gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta 3901 ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc 3961 acaatatttc aagctatacc aagcatacaa tcaactatct catatacaat gtctctcgag 4021 atgcccaaaa agaaaagaaa agtgggtagt atggcttcta actttactca gttcgttctc 4081 gtcgacaatg gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacgggatc 4141 gctgaatgga tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt 4201 cagagctctg cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa aggcgcctgg 4261 cgttcgtact taaatatgga actaaccatt ccaattttcg ccacgaattc cgactgcgag 4321 cttattgtta aggcaatgca aggtctccta aaagatggaa acccgattcc ctcagcaatc 4381 gcagcaaact ccggcatcta cggctccgga cgagccgatg ctctcgacga ttttgacctc 4441 gatatgctgg gatccgacgc tctcgatgac ttcgaccttg acatgctggg gagtgacgct 4501 ctggacgact tcgatctcga catgctgggc tccgacgccc tggatgactt cgatctggac 4561 atgctcatca actgaagtgc ttttaactaa gaattattag tcttttctgc ttattttttc 4621 atcatagttt agaacacttt atattaacga atagtttatg aatctattta ggtttaaaaa 4681 ttgatacagt tttataagtt actttttcaa agactcgtgc tgtctattgc ataatgcact 4741 ggaaggggaa aaaaaaggtg cacacgcgtg gctttttctt gaatttgcag tttgaaaaat 4801 aactacatgg atgataagaa aacatggagt acagtcactt tgagaacctt caatcagctg 4861 gtaacgtctt cgttaattgg atactcaaaa aagatggata gcatgaatca caagatggaa 4921 ggaaatgcgg gccacgacca cagtgatatg catatgggag atggagatga tacctggccc 4981 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt 5041 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt 5101 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga 5161 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag 5221 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt 5281 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac 5341 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc 5401 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa 5461 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt 5521 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat 5581 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata 5641 aacacacata aacaaacaaa atgctcgagg acaagaagta ttctatcgga ctggccatcg 5701 ggactaatag cgtcgggtgg gccgtgatca ctgacgagta caaggtgccc tctaagaagt 5761 tcaaggtgct cgggaacacc gaccggcatt ccatcaagaa aaatctgatc ggagctctcc 5821 tctttgattc aggggagacc gctgaagcaa cccgcctcaa gcggactgct agacggcggt 5881 acaccaggag gaagaaccgg atttgttacc ttcaagagat attctccaac gaaatggcaa 5941 aggtcgacga cagcttcttc cataggctgg aagaatcatt cctcgtggaa gaggataaga 6001 agcatgaacg gcatcccatc ttcggtaata tcgtcgacga ggtggcctat cacgagaaat 6061 acccaaccat ctaccatctt cgcaaaaagc tggtggactc aaccgacaag gcagacctcc 6121 ggcttatcta cctggccctg gcccacatga tcaagttcag aggccacttc ctgatcgagg 6181 gcgacctcaa tcctgacaat agcgatgtgg ataaactgtt catccagctg gtgcagactt 6241 acaaccagct ctttgaagag aaccccatca atgcaagcgg agtcgatgcc aaggccattc 6301 tgtcagcccg gctgtcaaag agccgcagac ttgagaatct tatcgctcag ctgccgggtg 6361 aaaagaaaaa tggactgttc gggaacctga ttgctctttc acttgggctg actcccaatt 6421 tcaagtctaa tttcgacctg gcagaggatg ccaagctgca actgtccaag gacacctatg 6481 atgacgatct cgacaacctc ctggcccaga tcggtgacca atacgccgac cttttccttg 6541 ctgctaagaa tctttctgac gccatcctgc tgtctgacat tctccgcgtg aacactgaaa 6601 tcaccaaggc ccctctttca gcttcaatga ttaagcggta tgatgagcac caccaggacc 6661 tgaccctgct taaggcactc gtccggcagc agcttccgga gaagtacaag gaaatcttct 6721 ttgaccagtc aaagaatgga tacgccggct acatcgacgg aggtgcctcc caagaggaat 6781 tttataagtt tatcaaacct atccttgaga agatggacgg caccgaagag ctcctcgtga 6841 aactgaatcg ggaggatctg ctgcggaagc agcgcacttt cgacaatggg agcattcccc 6901 accagatcca tcttggggag cttcacgcca tccttcggcg ccaagaggac ttctacccct 6961 ttcttaagga caacagggag aagattgaga aaattctcac tttccgcatc ccctactacg 7021 tgggacccct cgccagagga aatagccggt ttgcttggat gaccagaaag tcagaagaaa 7081 ctatcactcc ctggaacttc gaagaggtgg tggacaaggg agccagcgct cagtcattca 7141 tcgaacggat gactaacttc gataagaacc tccccaatga gaaggtcctg ccgaaacatt 7201 ccctgctcta cgagtacttt accgtgtaca acgagctgac caaggtgaaa tatgtcaccg 7261 aagggatgag gaagcccgca ttcctgtcag gcgaacaaaa gaaggcaatt gtggaccttc 7321 tgttcaagac caatagaaag gtgaccgtga agcagctgaa ggaggactat ttcaagaaaa 7381 ttgaatgctt cgactctgtg gagattagcg gggtcgaaga tcggttcaac gcaagcctgg 7441 gtacctacca tgatctgctt aagatcatca aggacaagga ttttctggac aatgaggaga 7501 acgaggacat ccttgaggac attgtcctga ctctcactct gttcgaggac cgggaaatga 7561 tcgaggagag gcttaagacc tacgcccatc tgttcgacga taaagtgatg aagcaactta 7621 aacggagaag atataccgga tggggacgcc ttagccgcaa actcatcaac ggaatccggg 7681 acaaacagag cggaaagacc attcttgatt tccttaagag cgacggattc gctaatcgca 7741 acttcatgca acttatccat gatgattccc tgacctttaa ggaggacatc cagaaggccc 7801 aagtgtctgg acaaggtgac tcactgcacg agcatatcgc aaatctggct ggttcacccg 7861 ctattaagaa gggtattctc cagaccgtga aagtcgtgga cgagctggtc aaggtgatgg 7921 gtcgccataa accagagaac attgtcatcg agatggccag ggaaaaccag actacccaga 7981 agggacagaa gaacagcagg gagcggatga aaagaattga ggaagggatt aaggagctcg 8041 ggtcacagat ccttaaagag cacccggtgg aaaacaccca gcttcagaat gagaagctct 8101 atctgtacta ccttcaaaat ggacgcgata tgtatgtgga ccaagagctt gatatcaaca 8161 ggctctcaga ctacgacgtg gacgccatcg tccctcagag cttcctcaaa gacgactcaa 8221 ttgacaataa ggtgctgact cgctcagaca agaaccgggg aaagtcagat aacgtgccct 8281 cagaggaagt cgtgaaaaag atgaagaact attggcgcca gcttctgaac gcaaagctga 8341 tcactcagcg gaagttcgac aatctcacta aggctgagag gggcggactg agcgaactgg 8401 acaaagcagg attcattaaa cggcaacttg tggagactcg gcagattact aaacatgtcg 8461 cccaaatcct tgactcacgc atgaatacca agtacgacga aaacgacaaa cttatccgcg 8521 aggtgaaggt gattaccctg aagtccaagc tggtcagcga tttcagaaag gactttcaat 8581 tctacaaagt gcgggagatc aataactatc atcatgctca tgacgcatat ctgaatgccg 8641 tggtgggaac cgccctgatc aagaagtacc caaagctgga aagcgagttc gtgtacggag 8701 actacaaggt ctacgacgtg cgcaagatga ttgccaaatc tgagcaggag atcggaaagg 8761 ccaccgcaaa gtacttcttc tacagcaaca tcatgaattt cttcaagacc gaaatcaccc 8821 ttgcaaacgg tgagatccgg aagaggccgc tcatcgagac taatggggag actggcgaaa 8881 tcgtgtggga caagggcaga gatttcgcta ccgtgcgcaa agtgctttct atgcctcaag 8941 tgaacatcgt gaagaaaacc gaggtgcaaa ccggaggctt ttctaaggaa tcaatcctcc 9001 ccaagcgcaa ctccgacaag ctcattgcaa ggaagaagga ttgggaccct aagaagtacg 9061 gcggattcga ttcaccaact gtggcttatt ctgtcctggt cgtggctaag gtggaaaaag 9121 gaaagtctaa gaagctcaag agcgtgaagg aactgctggg tatcaccatt atggagcgca 9181 gctccttcga gaagaaccca attgactttc tcgaagccaa aggttacaag gaagtcaaga 9241 aggaccttat catcaagctc ccaaagtata gcctgttcga actggagaat gggcggaagc 9301 ggatgctcgc ctccgctggc gaacttcaga agggtaatga gctggctctc ccctccaagt 9361 acgtgaattt cctctacctt gcaagccatt acgagaagct gaaggggagc cccgaggaca 9421 acgagcaaaa gcaactgttt gtggagcagc ataagcatta tctggacgag atcattgagc 9481 agatttccga gttttctaaa cgcgtcattc tcgctgatgc caacctcgat aaagtcctta 9541 gcgcatacaa taagcacaga gacaaaccaa ttcgggagca ggctgagaat atcatccacc 9601 tgttcaccct caccaatctt ggtgcccctg ccgcattcaa gtacttcgac accaccatcg 9661 accggaaacg ctatacctcc accaaagaag tgctggacgc caccctcatc caccagagca 9721 tcaccggact ttacgaaact cggattgacc tctcacagct cggaggggat gagggagctg 9781 atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtagat ccaaaaaaga 9841 agagaaaggt atagaattct accggcggcc gccaccgcgg tggagctcta agcaaatagc 9901 taaattatat acgaattaat attatgatta agtgtttacg tgagtgcgat atttttatta 9961 ctatcttata cagttgtata tactctataa aatgagttgt ctattaatta acgcgatgaa 10021 tgctttctgg gtttacctct ccaacaactc tagtttactt ctcaatacat tcaattgtat 10081 ttgatttgtc aatacttcat cattaatcaa ttctatagtt ttgtttttct cgtttatttc 10141 caaatttaat gcatcaattt tattattcaa tttgtcgttg attttggtta atgattttat 10201 ggtttgatct ctggcattga ttgtttgtgt tagtttttca ttattgataa ttaaattatt 10261 taagttagtt atcaactcgg tgttttcaag tttcaagttt tcaatttctt tagagtttat 10321 tagatttgtc aaagtttctg aattgcttga ttggtcctgt agaagagtat ttgttgttgt 10381 ggataattga ttcaattttt gagacaattg ctggaaggcg ttgaaatatc tagcatcaat 10441 ctcatggttt ttttcccgag agtctcgtag attcaattgt tttaatatat cttgggacca 10501 ctcttgattt gaactcatgg aaattaaact gggtgttgtg ttgtggtgta atgattgtac 10561 cccctttgct tataattgtg tggccaccga agtcggtgat gctgtcgccg aagaagttaa 10621 gaaaatcctt gcttaaaaag attctctttt tttatgatat ttgtacataa actttataaa 10681 tgaaattcat aatagaaacg acacgaaatt acaaaatgga atatgttcat agggtagacg 10741 aaactatata cgcaatctac atacatttat caagaaggag aaaaaggagg atgtaaagga 10801 atacaggtaa gcaaattgat actaatggct caacgtgata aggaaaaaga attgcacttt 10861 aacattaata ttgacaagga ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca 10921 tgaaactatg attcctaatt tatatattgg aggattttct ctaaaaaaaa aaaaatacaa 10981 caaataaaaa acactcaatg acctgaccat ttgatggagt ttaagtcaat accttcttga 11041 accatttccc ataatggtga aagttccctc aagaatttta ctctgtcaga aacggcctta 11101 acgacgtagg ccggccattt aaatgtttaa acggccacgc tggcccaatt cgccctatag 11161 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg 11221 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga 11281 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg 11341 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 11401 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 11461 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 11521 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 11581 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 11641 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 11701 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 11761 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc 11821 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt 11881 tgtgagttta gtattaggat ctctaccctt ggcgaaaagt cctctgccaa caatgatgat 11941 atctgatcca ccacttacaa cttcgtcgac ggttctgtac tgctgaccca atatgcgtat 12001 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga 12061 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt 12121 gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 12181 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc 12241 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca 12301 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc 12361 atgataataa tggtttctta gacgtcggcg cgccgaggac gtcaggtggc acttttcggg 12421 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 12481 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 12541 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 12601 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 12661 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 12721 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 12781 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 12841 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 12901 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 12961 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 13021 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 13081 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 13141 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 13201 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 13261 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 13321 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 13381 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 13441 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 13501 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 13561 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 13621 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 13681 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 13741 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 13801 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 13861 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 13921 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 13981 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 14041 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 14101 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 14161 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 14221 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 14281 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 14341 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca 14401 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 14461 attaggcacc ccaggcttta cactttatgc ttccggctcc tatgttgtgt ggaattgtga 14521 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcggaatta 14581 accctcacta aagggaacaa aagctg //