LOCUS Exported 10497 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses OsTIR1 via the ADH1 promoter, and the mGFP-AID via the TEF1 promoter in yeast. Both cassettes are integrated in HO locus. Assembly and characterization of the auxin-inducible degradation.. ACCESSION . VERSION . KEYWORDS pG2J SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10497) AUTHORS Papagiannakis A, Niebel B, Wit EC, Heinemann M TITLE Autonomous Metabolic Oscillations Robustly Gate the Early and Late Cell Cycle. JOURNAL Mol Cell. 2017 Jan 19;65(2):285-295. doi: 10.1016/j.molcel.2016.11.018. Epub 2016 Dec 15. PUBMED 27989441 REFERENCE 2 (bases 1 to 10497) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10497 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1..22 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." protein_bind 77..93 /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 82..104 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 101..117 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 101..117 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 1382..1778 /gene="S. cerevisiae ADH1" /label=ADH1 promoter /note="promoter for alcohol dehydrogenase 1" CDS 1797..3521 /codon_start=1 /product="rice transport inhibitor response 1 protein" /label=OsTIR1 /note="F-box protein that mediates auxin-dependent interaction of a ubiquitin ligase with an auxin-inducible degron (Tan et al., 2007)" /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL" CDS 3549..3578 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3585..3614 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3621..3650 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" terminator 3800..3965 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(3855..3879) /label=pHybLex-R /note="ADH terminator, reverse primer" terminator 3982..4171 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" CDS complement(4393..5109) /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4435..4456) /label=EGFP-C /note="EGFP, forward primer" primer_bind 4784..4803 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5043..5064 /label=EGFP-N /note="EGFP, reverse primer" promoter complement(5127..5525) /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" gene 5608..6964 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 5608..5951 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 5952..6761 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(6019..6038) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 6767..6964 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(7572..7591) /label=T7 /note="T7 promoter, forward primer" promoter complement(7573..7591) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7598..7615) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7598..7614) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7607..7629) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(7914..7933) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8046..8067 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(8221..8240) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8340..8362 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8400..8418) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8486..8590 /gene="bla" /label=AmpR promoter CDS 8591..9451 /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(8809..8828) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9622..10210 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10111..10130 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10364..10381 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccgcggct 61 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 121 gattacgcca ctagtccgag gcctcgagat ccgatatcgc cgtggcggcc gccagctgaa 181 gcttaattat cctgggcacg agtgaaacaa agctaaaacc tttatttagc atggccattg 241 aatgtaacaa ttatatatat cgcaagcaca aaaaatcaag gagagagaac taccactttg 301 ttcatgtgta caatgttcat tatctccata agcaaaaaaa aaaaaataga aaacatatgc 361 tataaggttg atattctcac gagtaagcgg cacttgctac ttattgacat tgcagatttt 421 tggctacaga aatagtatat tagagattat aattgctaat caaatcaaaa tataaaatta 481 gtaaaccaaa ccatttatac ccttccttag tagttatgga ttgtttttta atgatatttc 541 tgcaaaccaa agaaagattg ttatccagat agaatttagt tttgatattc atttttttgt 601 tgaagattga acgccatatc tgggcctcat aattcaaaag acggtgccat tatcggtagc 661 gtttcgcatt gtactggatt tcagaaattt cacagttgat gaatcgaaaa gaatggtctc 721 attgcaacac gtaaggttaa gatgtccctt tttaccatta taggcaataa atgaatcata 781 aaacgaccgt atactggtga aatagtaggg agaacgagta cctgtagtaa aaagtataaa 841 tcatagttaa tcgggcaatg tccctcgatc aaggagtatt gtgtcatgtt cgagacaaac 901 gccaacattt ttgtttcttt tggacaaatg ttgtttgcat ttatgatccg ttatattttg 961 atctaatgta gagttgcacg tagttcttac tggcaaagaa atcgatgcat accaaaaaag 1021 aataaaggtg atatttgatc tttaccgttt agttccaacg taaaattgtg cctttggact 1081 taaaatggcg gggtgtacaa tatggacttc ctcttttctg gcaaccaaac ccatacatcg 1141 ggattcctat aataccttcg ttggtctccc taacatgtag gtggcggagg ggagatatac 1201 aatagaacag ataccagaca agacataatg ggctaaacaa gactacacca attacactgc 1261 ctcattgatg gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc 1321 atatcgaagt ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat 1381 gcaacttctt ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc 1441 aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa 1501 cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt 1561 ccttcattca cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac 1621 ttgaatttga aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgcaat 1681 tattaatctt ttgtttcctc gtcattgttc tcgttccctt tcttccttgt ttctttttct 1741 gcacaatatt tcaagctata ccaagcatac aatcaactat ctcatataca actagaatga 1801 cgtacttccc ggaggaggtg gtggagcaca tcttcagctt cctgccggcg cagcgcgacc 1861 gcaacacggt ctcgctcgtc tgcaaggtgt ggtacgagat cgagaggctg agccgccgcg 1921 gcgtcttcgt gggcaactgc tacgccgtgc gcgccggccg cgtcgccgcg cggttcccca 1981 acgtgcgggc gctcacggtg aaggggaagc cccacttcgc cgacttcaac ctcgtgcccc 2041 ccgactgggg cggctacgcg gggccgtgga tcgaggcggc cgcgagggga tgccacggcc 2101 tggaggagct caggatgaag cggatggtgg tgtccgacga gagcctcgag ctgctggctc 2161 gctcgttccc gcggttcagg gctcttgttc ttatcagctg cgaggggttc agcactgacg 2221 ggctagccgc cgtcgcgagc cattgcaagc ttctgaggga gttggatttg caggaaaatg 2281 aagtggagga tcgagggcct aggtggcttt cctgcttccc tgattcctgc acatcacttg 2341 tctcattgaa ttttgcctgc atcaaagggg aggttaatgc tggttcactg gagagacttg 2401 ttagcaggtc cccaaacctg cggagtttga ggctgaatcg atctgtatcg gtagatacac 2461 ttgcaaagat actactgcgt acccctaact tggaggattt ggggacaggg aatttgacag 2521 atgacttcca aactgagtcc tactttaagc ttaccagtgc tctggagaaa tgcaagatgt 2581 tgaggagttt gtctggattc tgggatgctt ctcctgtttg cctgtcattt atctaccccc 2641 tgtgtgctca actgacagga ttgaacttga gctatgcacc cacacttgat gcttctgacc 2701 ttacaaaaat gattagccgc tgtgtgaagc tccaacgcct ttgggtactg gattgtatct 2761 cggacaaagg cttgcaagtg gtggcctcca gttgcaaaga cttgcaagaa ctcagggtat 2821 ttccatcaga tttctacgta gctggttatt ctgcagtgac agaggaggga cttgttgcag 2881 tatccttggg ctgtccaaaa ctgaactcac tactgtactt ctgtcaccaa atgactaatg 2941 ctgcactagt tactgtcgcc aagaactgtc caaatttcac acgattcaga ctttgtattc 3001 ttgagccagg gaagcctgat gttgtgacaa gccaaccatt agatgaaggc tttggagcta 3061 ttgttcgtga gtgcaaggga ttacaacgtt tgtcaatatc tggtcttctc acagacaaag 3121 ttttcatgta tattgggaaa tatgcaaaac aacttgagat gctttctata gcatttgctg 3181 gtgacagtga taagggtatg atgcatgtta tgaatggatg caagaattta aggaaactgg 3241 agataagaga tagcccgttt ggtgatgctg cactcttggg gaattttgct aggtacgaga 3301 caatgcgatc cctttggatg tcatcttgca atgtcacgtt aaaggggtgc caagtccttg 3361 cgtcaaagat gccgatgctc aatgttgagg tcataaatga gcgggatggt agcaatgaaa 3421 tggaggaaaa ccatggagat ctgcctaaag tggagaaatt atatgtgtac cgcacaactg 3481 ctggggcgag ggatgatgca ccaaattttg ttaaaatcct agtcgactcc ggttctgctg 3541 ctagtggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa aagctaatct 3601 ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg aacggatcca 3661 ctagctaaaa caattcttcg ccagaggttt ggtcaagtct ccaatcaagg ttgtcggctt 3721 gtctaccttg ccagaaattt acgaaaagat ggaaaagggt caaatcgttg gtagatacgt 3781 tgttgacact tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt 3841 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt 3901 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt 3961 cgctcgtacg ctgagctgga tcttcgagcg tcccaaaacc ttctcaagca aggttttcag 4021 tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa 4081 taacgttctt aatactaaca taactataaa aaaataaata gggacctaga cttcaggttg 4141 tctaactcct tccttttcgg ttagagcgga tcttagctag ccgcggtacc aagcttactc 4201 gagttacccg ggtgatacct tcacgaacgc cgccgcctcc gggccaccgc ttgatttttg 4261 gcaggaaacc atcacgttct tccggtatga tctcaccggt ggccatccca caacttgtgc 4321 cttggccgga ggtttggctg gatctttagg ggtacctgcc ccagcaccag ccccggcgcc 4381 tgcaccgtcg accttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa 4441 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactgggt 4501 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg 4561 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac 4621 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta 4681 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat 4741 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc 4801 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa 4861 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt 4921 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt 4981 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt 5041 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt 5101 gctcaccatt ttaataacct aggaaactta gattagattg ctatgctttc tttctaatga 5161 gcaagaagta aaaaaagttg taatagaaca agaaaaatga aactgaaact tgagaaattg 5221 aagaccgttt attaacttaa atatcaatgg gaggtcatcg aaagagaaaa aaatcaaaaa 5281 aaaaattttc aagaaaaaga aacgtgataa aaatttttat tgcctttttc gacgaagaaa 5341 aagaaacgag gcggtctctt ttttcttttc caaaccttta gtacgggtaa ttaacgacac 5401 cctagaggaa gaaagagggg aaatttagta tgctgtgctt gggtgttttg aagtggtacg 5461 gcgatgcgcg gagtccgaga aaatctggaa gagtaaaaaa ggagtagaaa cattttgaag 5521 ctatggtgtg gaattccagc tgcgggttaa ttaaggcgcg ccagatctgt ttagcttgcc 5581 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt 5641 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat 5701 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat 5761 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat 5821 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc 5881 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa 5941 cataaacaac catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca 6001 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga 6061 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag 6121 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta 6181 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca 6241 ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa 6301 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt 6361 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg 6421 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct 6481 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt 6541 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac 6601 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt 6661 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga 6721 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa 6781 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt 6841 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc 6901 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat 6961 actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacgagct cgaattaatt 7021 cctgggggaa caacttcaca gaatgttttg tcatattgtc gaagtggtca caaaacaaga 7081 gaagttccgc caattataaa aagggaaccc gtatatttca gcttcacgga tgatttccag 7141 ggtgagagta ctgtatatgg gcttacgata gaaggccata aaaatttctt gcttggcaac 7201 aaaatagaag tgaaatcatg tcgaggctgc tgtgtgggag aacagcataa aatatcacaa 7261 aaaaagaatc taaaacactg tgttgcttgt cccagaaagg gaatcaagta tttttataaa 7321 gattggagtg gtaaaaatcg agtatgtgct agatgctatg gaagatacaa attcagcggt 7381 catcactgta taaattgcaa gtatgtacca gaagcacgtg aagtgaaaaa ggcaaaagac 7441 aaaggcgaaa aattgggcat tacgcccgaa ggtttgccag ttaaaggacc agagtgtata 7501 aaatgtggcg gaatcttaca gtggcctatg cggccgctct agctagaact agtggatcga 7561 tccccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc 7621 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 7681 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 7741 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 7801 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg 7861 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt 7921 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 7981 gggtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 8041 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt 8101 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca 8161 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag 8221 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga 8281 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 8341 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg 8401 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc 8461 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 8521 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 8581 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 8641 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 8701 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 8761 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 8821 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 8881 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 8941 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 9001 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 9061 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 9121 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 9181 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 9241 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 9301 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 9361 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 9421 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 9481 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 9541 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 9601 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 9661 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 9721 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 9781 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 9841 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 9901 aagacgatag ttaccggata aggcacagcg gtcgggctga acggggggtt cgtgcacaca 9961 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 10021 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 10081 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 10141 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 10201 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 10261 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 10321 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 10381 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 10441 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaat //