LOCUS V020243 6917 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020243 VERSION V020243 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 683..1573 /label="TRIM52(NM_032765)" /note="TRIM52(NM_032765)" /gene="TRIM52" CDS 1580..1669 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1693..2281 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2347..2395 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2597..3025 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3039..3368 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3435..4229 /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 4405..4526 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4575..4591) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4599..4615 /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(4623..4653) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4668..4689 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4977..5565) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5736..6596) /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(6597..6701) /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 gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatggctgg ttatgccact actcccagcc ccatgcagac 721 ccttcaggag gaagcggtgt gtgccatctg cttggattac ttcaaggacc ccgtgtccat 781 cagctgtggg cacaacttct gccgagggtg tgtgacccag ctgtggagta aggaggacga 841 ggaggaccag aacgaggagg aagatgaatg ggaggaggag gaggacgagg aagcggtggg 901 ggccatggat ggatgggacg gctccattcg agaggtgttg tatcggggga atgctgacga 961 agagttgttc caagaccaag atgacgatga actctggctc ggtgacagtg gtataactaa 1021 ttgggacaac gtagactata tgtgggacga ggaggaagaa gaagaagagg aagatcagga 1081 ctattaccta ggaggcttga gacctgacct gagaattgat gtctaccgag aagaagaaat 1141 actggaagca tacgatgagg acgaagatga agagctgtat cctgacatcc acccgcctcc 1201 ttccttgccc cttccagggc agttcacctg cccccagtgc cgaaagagct ttacacgtcg 1261 cagctttcgt cccaacttgc agctggccaa catggtccag ataattcgcc agatgtgccc 1321 cactccttat cggggaaacc ggagtaatga tcagggcatg tgctttaaac accaggaagc 1381 cctgaaactc ttctgtgagg tggacaaaga ggccatctgt gtggtgtgcc gagaatccag 1441 gagccacaaa cagcacagcg tgctgccttt ggaggaggtg gtgcaggagt accaggaaat 1501 aaagttggaa acaactctgg tgggaatact tcagatagag caagaaagca ttcacagcaa 1561 ggcctataat cagctcgagg aacaaaaact aatatcagag gaagatttgg aacagaaact 1621 aatcagtgaa gaagatttag agcaaaagtt aatttccgag gaggacttat agtgagggcc 1681 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 1741 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 1801 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 1861 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 1921 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 1981 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 2041 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt 2101 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 2161 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 2221 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 2281 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg 2341 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa 2401 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 2461 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 2521 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc 2581 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2641 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2701 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 2761 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 2821 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 2881 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 2941 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 3001 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3121 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3181 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 3241 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3301 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 3361 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 3421 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3481 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3541 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3601 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3661 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3721 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3781 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3841 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3901 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 3961 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4021 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4081 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4141 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4201 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac 4261 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 4321 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 4381 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 4441 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4501 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 4561 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 4621 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 4681 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 4741 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 4921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 4981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5161 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 5401 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 5521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 5581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 5641 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 5701 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 5761 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 5821 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 5881 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 5941 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6001 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6061 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6121 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6181 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 6241 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 6301 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 6361 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6421 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 6481 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 6541 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 6601 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 6661 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6721 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa 6781 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 6841 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 6901 tgaagaatct gcttagg //