LOCUS Exported 6803 bp ds-DNA circular SYN 28-MAY-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBSWHHC_p53 SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 6803) AUTHORS Mireia TITLE Direct Submission JOURNAL Exported 23 Sep 2019 from SnapGene 2.6.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6803 /organism="recombinant plasmid" /mol_type="other DNA" promoter 30..734 /gene="S. cerevisiae ADH1" /note="ADH1 promoter" /note="promoter for alcohol dehydrogenase 1" CDS 762..1202 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="DNA binding domain of the GAL4 transcriptional activator" /note="GAL4 DNA binding domain" /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS" promoter 1213..1231 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 1245..1247 /codon_start=1 /product="start codon for in vitro transcription/translation" /note="ATG" /translation="M" CDS 1251..1280 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /translation="EQKLISEEDL" misc_feature 1281..1309 /label=MCS /note="MCS" /note="multiple cloning site" CDS 1310 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="activation domain of the GAL4 transcriptional activator" /note="GAL4 activation domain" /translation="" protein_bind 1317..1341 /gene="mutant version of attB" /bound_moiety="BP Clonase(TM)" /note="attB1" /note="recombination site for the Gateway(R) BP reaction" CDS 1344..2303 /codon_start=1 /note="p53" /translation="PVTETPGPVAPAPATPWPLSSFVPSQKTYQGNYGFHLGFLQSGTA KSVMCTYSPPLNKLFCQLAKTCPVQLWVSATPPAGSRVRAMAIYKKSQHMTEVVRRCPH HERCSDGDGLAPPQHLIRVEGNLYPEYLEDRQTFRHSVVVPYEPPEAGSEYTTIHYKYM CNSSCMGGMNRRPILTIITLEDSSGNLLGRDSFEVRVCACPGRDRRTEEENFRKKEVLC PELPPGSAKRALPTCTSASPPQKKKPLDGEYFTLKIRGRKRFEMFRELNEALELKDAHA TEESGDSRAHSSYLKTKKGQSTSRHKKTMVKKVGPDSD" protein_bind complement(2305..2329) /gene="mutant version of attB" /bound_moiety="BP Clonase(TM)" /note="attB2" /note="recombination site for the Gateway(R) BP reaction" misc_feature 2340..2363 /label=MCS /note="MCS" /note="multiple cloning site" terminator 2368..2415 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2442..2536 /gene="S. cerevisiae ADH1" /note="ADH1 terminator" /note="transcription terminator for alcohol dehydrogenase 1" misc_feature 2537..2596 /note="H1_overlap_site" misc_feature 2593..2643 /note="Self_overlap_site" rep_origin complement(2958..3546) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" misc_feature 3875..3922 /note="HSV TK poly(A) signal" /note="herpesvirus thymidine kinase polyadenylation signal" CDS complement(4154..4945) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /note="SmR" /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" promoter complement(4946..5050) /gene="bla" /note="AmpR promoter" misc_feature 5126..5176 /note="Self_overlap_site" misc_feature 5173..5232 /note="H2_overlap_site" promoter 5248..5529 /gene="S. cerevisiae TRP1" /note="TRP1 promoter" CDS 5530..6204 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /note="TRP1" /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" rep_origin complement(6307..6762) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg 61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg 121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac 181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga 241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt 301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct 361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac 421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga 481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc 541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt 601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca 661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa 721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta 781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga 841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt 901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc 961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac 1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg 1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa 1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat 1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc 1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcccgggg atctccacaa 1321 gtttgtacaa aaaagcaggc ttacctgtca ccgagacccc tgggccagtg gcccctgccc 1381 cagccactcc atggcccctg tcatcttttg tcccttctca aaaaacttac cagggcaact 1441 atggcttcca cctgggcttc ctgcagtctg ggacagccaa gtctgttatg tgcacgtact 1501 ctcctcccct caataagcta ttctgccagc tggcgaagac gtgccctgtg cagttgtggg 1561 tcagcgccac acctccagct gggagccgtg tccgcgccat ggccatctac aagaagtcac 1621 agcacatgac ggaggtcgtg agacgctgcc cccaccatga gcgctgctcc gatggtgatg 1681 gcctggctcc tccccagcat cttatccggg tggaaggaaa tttgtatccc gagtatctgg 1741 aagacaggca gacttttcgc cacagcgtgg tggtacctta tgagccaccc gaggccggct 1801 ctgagtatac caccatccac tacaagtaca tgtgtaatag ctcctgcatg gggggcatga 1861 accgccgacc tatccttacc atcatcacac tggaagactc cagtgggaac cttctgggac 1921 gggacagctt tgaggttcgt gtttgtgcct gccctgggag agaccgccgt acagaagaag 1981 aaaatttccg caaaaaggaa gtcctttgcc ctgaactgcc cccagggagc gcaaagagag 2041 cgctgcccac ctgcacaagc gcctctcccc cgcaaaagaa aaaaccactt gatggagagt 2101 atttcaccct caagatccgc gggcgtaaac gcttcgagat gttccgggag ctgaatgagg 2161 ccttagagtt aaaggatgcc catgctacag aggagtctgg agacagcagg gctcactcca 2221 gctacctgaa gaccaagaag ggccagtcta cttcccgcca taaaaaaaca atggtcaaga 2281 aagtggggcc tgactcagac tgaaacccag ctttcttgta caaagtggta agtaactaag 2341 atccgtcgac ctgcagcggc cgcataacta gcataacccc ttggggcctc taaacgggtc 2401 ttgaggggtt ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc ttatgattta 2461 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact 2521 cttaggtttt aaaacgttat ccctagttta aacacccata atacccataa tagctgtttg 2581 ccaattaccc tgttatccct attacttcta aatctgagga ggaaaatcga atcgctacct 2641 tagcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg 2701 ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat taggtgttac 2761 atgctgttca tctgttacat tgtcgatctg ttcatggtga acaggcgctc ggtcgttcgg 2821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 2881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 2941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 3001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 3061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 3121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 3181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 3301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 3361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 3421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 3481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 3541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 3601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 3661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg 3721 cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt 3781 ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca 3841 gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt 3901 ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt 3961 gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg 4021 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg 4081 gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac 4141 cccagagtcc cgcttatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa 4201 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct 4261 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg 4321 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg 4381 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg 4441 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg 4501 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa 4561 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa 4621 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg 4681 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt 4741 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat 4801 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca 4861 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata 4921 cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga agcatttatc 4981 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 5041 gggttccgcg cacatctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 5101 tatcacgagg ccctttcgtc ttcacttatc cctattactt ctaaatctga ggaggaaaat 5161 cgaatcgcta ccttaggacc gttatagtta ctatcaccat atatgtaact ggtgattctt 5221 tcgctacctt agagtgcacc atagatcaac gacattacta tatatataat ataggaagca 5281 tttaatagaa cagcatcgta atatatgtgt actttgcagt tatgacgcca gatggcagta 5341 gtggaagata ttctttattg aaaaatagct tgtcacctta cgtacaatct tgatccggag 5401 cttttctttt tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg agagggccaa 5461 gagggagggc attggtgact attgagcacg tgagtatacg tgattaagca cacaaaggca 5521 gcttggagta tgtctgttat taatttcaca ggtagttctg gtccattggt gaaagtttgc 5581 ggcttgcaga gcacagaggc cgcagaatgt gctctagatt ccgatgctga cttgctgggt 5641 attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag gaaaatttca 5701 agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt tggcgtgttt 5761 cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat tgatatcgtc 5821 caactgcatg gagatgagtc gtggcaagaa taccaagagt tcctcggttt gccagttatt 5881 aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc acagaaacct 5941 cattcgttta ttcccttgtt tgattcagaa gcaggtggga caggtgaact tttggattgg 6001 aactcgattt ctgactgggt tggaaggcaa gagagccccg aaagcttaca ttttatgtta 6061 gctggtggac tgacgccaga aaatgttggt gatgcgctta gattaaatgg cgttattggt 6121 gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa aatagcaaat 6181 ttcgtcaaaa atgctaagaa ataggttatt actgagtagt atttatttaa gtattgtttg 6241 tgcacttgcc gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 6301 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 6361 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 6421 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 6481 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 6541 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 6601 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 6661 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 6721 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct 6781 gcgcaactgt tgggaagggc gat //