LOCUS Exported 9972 bp ds-DNA circular 05-JUL-2019 COMMENT Created by Takara Bio USA, Inc. http://www.takarabio.com COMMENT Cat. Nos. 630466, 630489, 630498, 630499 SOURCE synthetic DNA construct FEATURES Location/Qualifiers source 1..9972 rep_origin 368..1532 /direction=RIGHT /label=2u ori promoter 2306..3010 /gene="S. cerevisiae ADH1" /label=ADH1 promoter CDS 3056..3076 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS CDS 3092..3433 /label=GAL4 activation domain regulatory 3464..3473 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3476..3502 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA CDS 3647..3667 /label=SV40 NLS misc_feature 3730..5395 /label=SV40 Large-T Antigen polyA_signal 5421..5542 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" terminator 5935..6122 /gene="S. cerevisiae ADH1" /label=ADH1 terminator CDS complement(6239..7333) /label=LEU2 /note="yeast auxotrophic marker" promoter complement(7334..7739) /gene="S. cerevisiae LEU2" /label=LEU2 promoter rep_origin 8334..8922 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(9093..9953) /label=AmpR promoter complement(join(9954..9972,1..86)) /gene="bla" /label=AmpR promoter ORIGIN 1 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 61 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa 121 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 181 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 241 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 301 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 361 caccataacg catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat 421 acaggcaaca cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg 481 ttgcattttc ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct 541 attctctagc tagaaagtat aggaacttca gagcgctttt gaaaaccaaa agcgctctga 601 agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta aaatattgcg 661 aataccgctt ccacaaacat tgctcaaaag tatctctttg ctatatatct ctgtgctata 721 tccctatata acctacccat ccacctttcg ctccttgaac ttgcatctaa actcgacctc 781 tacatttttt atgtttatct ctagtattac tctttagaca aaaaaattgt agtaagaact 841 attcatagag tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac gtagtatata 901 gagacaaaat agaagaaacc gttcataatt ttctgaccaa tgaagaatca tcaacgctat 961 cactttctgt tcacaaagta tgcgcaatcc acatcggtat agaatataat cggggatgcc 1021 tttatcttga aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag 1081 tcaggctttt tttatggaag agaaaataga caccaaagta gccttcttct aaccttaacg 1141 gacctacagt gcaaaaagtt atcaagagac tgcattatag agcgcacaaa ggagaaaaaa 1201 agtaatctaa gatgctttgt tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa 1261 aaaagaagta tagattcttt gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt 1321 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt 1381 tacaaaaatg aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt 1441 tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt 1501 tgttctacaa aatgaagcac agatgcttcg ttgcttgcat gcctgcaggt cgagatccgg 1561 gatcgaagaa atgatggtaa atgaaatagg aaatcaagga gcatgaaggc aaaagacaaa 1621 tataagggtc gaacgaaaaa taaagtgaaa agtgttgata tgatgtattt ggctttgcgg 1681 cgccgaaaaa acgagtttac gcaattgcac aatcatgctg actctgtggc ggacccgcgc 1741 tcttgccggc ccggcgataa cgctgggcgt gaggctgtgc ccggcggagt tttttgcgcc 1801 tgcattttcc aaggtttacc ctgcgctaag gggcgagatt ggagaagcaa taagaatgcc 1861 ggttggggtt gcgatgatga cgaccacgac aactggtgtc attatttaag ttgccgaaag 1921 aacctgagtg catttgcaac atgagtatac tagaagaatg agccaagact tgcgagacgc 1981 gagtttgccg gtggtgcgaa caatagagcg accatgacct tgaaggtgag acgcgcataa 2041 ccgctagagt actttgaaga ggaaacagca atagggttgc taccagtata aatagacagg 2101 tacatacaac actggaaatg gttgtctgtt tgagtacgct ttcaattcat ttgggtgtgc 2161 actttattat gttacaatat ggaagggaac tttacacttc tcctatgcac atatattaat 2221 taaagtccaa tgctagtaga gaaggggggt aacacccctc cgcgctcttt tccgattttt 2281 ttctaaaccg tggaatattt cggatatcct tttgttgttt ccgggtgtac aatatggact 2341 tcctcttttc tggcaaccaa acccatacat cgggattcct ataatacctt cgttggtctc 2401 cctaacatgt aggtggcgga ggggagatat acaatagaac agataccaga caagacataa 2461 tgggctaaac aagactacac caattacact gcctcattga tggtggtaca taacgaacta 2521 atactgtagc cctagacttg atagccatca tcatatcgaa gtttcactac cctttttcca 2581 tttgccatct attgaagtaa taataggcgc atgcaacttc ttttcttttt ttttcttttc 2641 tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg gaagacacta 2701 aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag agctgatgag 2761 gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact actctctaat 2821 gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa aaagtttgct 2881 gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc tcgtcattgt 2941 tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta taccaagcat 3001 acaatcaact ccaagctttg caaagatgga taaagcggaa ttaattcccg agcctccaaa 3061 aaagaagaga aaggtcgaat tgggtaccgc cgccaatttt aatcaaagtg ggaatattgc 3121 tgatagctca ttgtccttca ctttcactaa cagtagcaac ggtccgaacc tcataacaac 3181 tcaaacaaat tctcaagcgc tttcacaacc aattgcctcc tctaacgttc atgataactt 3241 catgaataat gaaatcacgg ctagtaaaat tgatgatggt aataattcaa aaccactgtc 3301 acctggttgg acggaccaaa ctgcgtataa cgcgtttgga atcactacag ggatgtttaa 3361 taccactaca atggatgatg tatataacta tctattcgat gatgaagata ccccaccaaa 3421 cccaaaaaaa gagatcttta atacgactca ctatagggcg agcgccgcca tggagtaccc 3481 atacgacgta ccagattacg ctcatatggc catggaggcc agtgaattcg gaactgatga 3541 atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc 3601 atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag 3661 aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt 3721 gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact 3781 gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta 3841 taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa 3901 ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata 3961 tttgatgtat agtgccttga ctagagatcc attttctgtt attgaggaaa gtttgccagg 4021 tgggttaaag gagcatgatt ttaatccaga agaagcagag gaaactaaac aagtgtcctg 4081 gaagcttgta acagagtatg caatggaaac aaaatgtgat gatgtgttgt tattgcttgg 4141 gatgtacttg gaatttcagt acagttttga aatgtgttta aaatgtatta aaaaagaaca 4201 gcccagccac tataagtacc atgaaaagca ttatgcaaat gctgctatat ttgctgacag 4261 caaaaaccaa aaaaccatat gccaacaggc tgttgatact gttttagcta aaaagcgggt 4321 tgatagccta caattaacta gagaacaaat gttaacaaac agatttaatg atcttttgga 4381 taggatggat ataatgtttg gttctacagg ctctgctgac atagaagaat ggatggctgg 4441 agttgcttgg ctacactgtt tgttgcccaa aatggattca gtggtgtatg actttttaaa 4501 atgcatggtg tacaacattc ctaaaaaaag atactggctg tttaaaggac caattgatag 4561 tggtaaaact acattagcag ctgctttgct tgaattatgt ggggggaaag ctttaaatgt 4621 taatttgccc ttggacaggc tgaactttga gctaggagta gctattgacc agtttttagt 4681 agtttttgag gatgtaaagg gcactggagg ggagtccaga gatttgcctt caggtcaggg 4741 aattaataac ctggacaatt taagggatta tttggatggc agtgttaagg taaacttaga 4801 aaagaaacac ctaaataaaa gaactcaaat atttccccct ggaatagtca ccatgaatga 4861 gtacagtgtg cctaaaacac tgcaggccag atttgtaaaa caaatagatt ttaggcccaa 4921 agattattta aagcattgcc tggaacgcag tgagtttttg ttagaaaaga gaataattca 4981 aagtggcatt gctttgcttc ttatgttaat ttggtacaga cctgtggctg agtttgctca 5041 aagtattcag agcagaattg tggagtggaa agagagattg gacaaagagt ttagtttgtc 5101 agtgtatcaa aaaatgaagt ttaatgtggc tatgggaatt ggagttttag attggctaag 5161 aaacagtgat gatgatgatg aagacagcca ggaaaatgct gataaaaatg aagatggtgg 5221 ggagaagaac atggaagact cagggcatga aacaggcatt gattcacagt cccaaggctc 5281 atttcaggcc cctcagtcct cacagtctgt tcatgatcat aatcagccat accacatttg 5341 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 5401 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5521 ccaaactcat caatgtatct tatcatgtct cgagctgcag atgaatcgta gatactgaaa 5581 aaccccgcaa gttcacttca actgtgcatc gtgcaccatc tcaatttctt tcatttatac 5641 atcgttttgc cttcttttat gtaactatac tcctctaagt ttcaatcttg gccatgtaac 5701 ctctgatcta tagaattttt taaatgacta gaattaatgc ccatcttttt tttggaccta 5761 aattcttcat gaaaatatat tacgagggct tattcagaag ctttggactt cttcgccaga 5821 ggtttggtca agtctccaat caaggttgtc ggcttgtcta ccttgccaga aatttacgaa 5881 aagatggaaa agggtcaaat cgttggtaga tacgttgttg acacttctaa ataagcgaat 5941 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa 6001 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg 6061 taggtcaggt tgctttctca ggtatagcat gaggtcgctc ttattgacca cacctctacc 6121 ggccggtcga aattccccta ccctatgaac atattccatt ttgtaatttc gtgtcgtttc 6181 tattatgaat ttcatttata aagtttatgt acaaatatca taaaaaaaga gaatcttttt 6241 aagcaaggat tttcttaact tcttcggcga cagcatcacc gacttcggtg gtactgttgg 6301 aaccacctaa atcaccagtt ctgatacctg catccaaaac ctttttaact gcatcttcaa 6361 tggccttacc ttcttcaggc aagttcaatg acaatttcaa catcattgca gcagacaaga 6421 tagtggcgat agggttgacc ttattctttg gcaaatctgg agcagaaccg tggcatggtt 6481 cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc caaggacgca gatggcaaca 6541 aacccaagga acctgggata acggaggctt catcggagat gatatcacca aacatgttgc 6601 tggtgattat aataccattt aggtgggttg ggttcttaac taggatcatg gcggcagaat 6661 caatcaattg atgttgaacc ttcaatgtag gaaattcgtt cttgatggtt tcctccacag 6721 tttttctcca taatcttgaa gaggccaaaa cattagcttt atccaaggac caaataggca 6781 atggtggctc atgttgtagg gccatgaaag cggccattct tgtgattctt tgcacttctg 6841 gaacggtgta ttgttcacta tcccaagcga caccatcacc atcgtcttcc tttctcttac 6901 caaagtaaat acctcccact aattctctga caacaacgaa gtcagtacct ttagcaaatt 6961 gtggcttgat tggagataag tctaaaagag agtcggatgc aaagttacat ggtcttaagt 7021 tggcgtacaa ttgaagttct ttacggattt ttagtaaacc ttgttcaggt ctaacactac 7081 ctgtacccca tttaggacca cccacagcac ctaacaaaac ggcatcagcc ttcttggagg 7141 cttccagcgc ctcatctgga agtgggacac ctgtagcttc gatagcagca ccaccaatta 7201 aatgattttc gaaatcgaac ttgacattgg aacgaacatc agaaatagct ttaagaacct 7261 taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct 7321 taggggcaga cattagaatg gtatatcctt gaaatatata tatatattgc tgaaatgtaa 7381 aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa aaacaatagg 7441 tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt 7501 ctctattcta tatgaaaagc cggttccggc gctctcacct ttcctttttc tcccaatttt 7561 tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa tttccagtca 7621 tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga ggaaaaaaat 7681 aatggttgct aagagattcg aactcttgca tcttacgata cctgagtatt cccacagttg 7741 ggggatctcg actctagcta gaggatcaat tcgtaatcat gtcatagctg tttcctgtgt 7801 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 7861 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 7921 tccagtcggg aaacctgtcg tgccagctga taacttcgta taatgtatgc tatacgaagt 7981 tattaggtct gaagaggagt ttacgtccag ccaagctagc ttggctgcag gtcgagcggc 8041 cgcgatccgg aacccttaat ataacttcgt ataatgtatg ctatacgaag ttatcagctg 8101 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 8161 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 8221 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 8281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 8341 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 8401 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 8461 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 8521 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 8581 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 8641 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 8701 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 8761 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 8821 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 8881 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 8941 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 9001 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 9061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 9121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 9181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 9241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 9301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 9361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 9421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 9481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 9541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 9601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 9661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 9721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 9781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 9841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 9901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 9961 ctttttcaat at //