LOCUS V027472 7280 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027472 VERSION V027472 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 775..1953 /codon_start=1 /gene="Homo sapiens TP53" /note="responds to cellular stresses to regulate the expression of target genes" /product="human tumor suppressor protein that is often mutated in cancers" /transl_table=1 /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD" /label="p53" CDS 1960..1986 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 2056..2644 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2710..2758 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2960..3388 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3402..3731 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3798..4592 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4768..4889 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4938..4954) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4962..4978 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4986..5016) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5031..5052 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5340..5928) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6099..6959) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6960..7064) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatggag 781 gagccgcagt cagatcctag cgtcgagccc cctctgagtc aggaaacatt ttcagaccta 841 tggaaactac ttcctgaaaa caacgttctg tcccccttgc cgtcccaagc aatggatgat 901 ttgatgctgt ccccggacga tattgaacaa tggttcactg aagacccagg tccagatgaa 961 gctcccagaa tgccagaggc tgctcccccc gtggcccctg caccagcagc tcctacaccg 1021 gcggcccctg caccagcccc ctcctggccc ctgtcatctt ctgtcccttc ccagaaaacc 1081 taccagggca gctacggttt ccgtctgggc ttcttgcatt ctgggacagc caagtctgtg 1141 acttgcacgt actcccctgc cctcaacaag atgttttgcc aactggccaa gacctgccct 1201 gtgcagctgt gggttgattc cacacccccg cccggcaccc gcgtccgcgc catggccatc 1261 tacaagcagt cacagcacat gacggaggtt gtgaggcgct gcccccacca tgagcgctgc 1321 tcagatagcg atggtctggc ccctcctcag catcttatcc gagtggaagg aaatttgcgt 1381 gtggagtatt tggatgacag aaacactttt cgacatagtg tggtggtgcc ctatgagccg 1441 cctgaggttg gctctgactg taccaccatc cactacaact acatgtgtaa cagttcctgc 1501 atgggcggca tgaaccggag gcccatcctc accatcatca cactggaaga ctccagtggt 1561 aatctactgg gacggaacag ctttgaggtg cgtgtttgtg cctgtcctgg gagagaccgg 1621 cgcacagagg aagagaatct ccgcaagaaa ggggagcctc accacgagct gcccccaggg 1681 agcactaagc gagcactgcc caacaacacc agctcctctc cccagccaaa gaagaaacca 1741 ctggatggag aatatttcac ccttcagatc cgtgggcgtg agcgcttcga gatgttccga 1801 gagctgaatg aggccttgga actcaaggat gcccaggctg ggaaggagcc aggggggagc 1861 agggctcact ccagccacct gaagtccaaa aagggtcagt ctacctcccg ccataaaaaa 1921 ctcatgttca agacagaagg gcctgactca gacgaattct acccatacga cgtaccagat 1981 tacgcttaac tcgagtctag agggcccctt aagggttcga tccctaccgg ttagtaatga 2041 gtttgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 2101 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 2161 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 2221 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 2281 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 2341 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 2401 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 2461 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 2521 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 2581 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 2641 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 2701 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat 2761 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 2821 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 2881 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg 2941 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 3001 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 3061 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc 3121 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 3181 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 3241 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 3301 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag 3361 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 3421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3481 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 3601 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 3661 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 3721 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 3781 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 3841 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 3901 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 3961 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 4021 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 4081 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 4141 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 4201 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 4261 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 4321 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 4381 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 4441 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 4501 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 4561 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc 4621 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 4681 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 4741 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 4801 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 4861 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 4921 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 4981 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 5041 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 5101 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 5161 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 5221 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 5281 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 5341 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 5401 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 5461 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 5521 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 5581 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 5641 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 5701 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 5761 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 5821 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 5881 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 5941 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 6001 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 6061 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 6121 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 6181 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 6241 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 6301 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 6361 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 6421 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 6481 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 6541 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 6601 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 6661 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 6721 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 6781 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 6841 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 6901 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 6961 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 7021 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 7081 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta 7141 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg 7201 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 7261 gcatgaagaa tctgcttagg //