LOCUS V034249 7510 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034249 VERSION V034249 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 706..2166 /label="TRIM50(NM_178125)" /note="TRIM50(NM_178125)" /gene="TRIM50" CDS 2173..2262 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2286..2874 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2940..2988 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3190..3618 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3632..3961 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4028..4822 /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 4998..5119 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5168..5184) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5192..5208 /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(5216..5246) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5261..5282 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5570..6158) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6329..7189) /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(7190..7294) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc ttggcaggtg 721 agcctgccag agctggagga ccggcttcag tgtcccatct gcctggaggt cttcaaggag 781 cccctgatgc tgcagtgtgg ccactcttac tgcaagggct gcctggtttc cctgtcctgc 841 cacctggatg ccgagctgcg ctgccccgtg tgccggcagg cggtggacgg cagcagctcc 901 ctgcccaacg tctccctggc cagggtgatc gaagccctga ggctccctgg ggacccggag 961 cccaaggtct gcgtgcacca ccggaacccg ctcagccttt tctgcgagaa ggaccaggag 1021 ctcatctgtg gcctctgcgg tctgctgggc tcccaccaac accacccggt cacgcccgtc 1081 tccaccgtct acagccgcat gaaggaggag ctcgcagccc tcatctctga gctgaagcag 1141 gagcagaaaa aggtggatga gctcatcgcc aaactggtga acaaccggac ccgaatcgtc 1201 aatgagtcgg atgtcttcag ctgggtgatc cgccgcgagt tccaggagct gcaccacctg 1261 gtggatgagg agaaggcccg ctgcctggag gggatagggg gtcacacccg tggcctggtg 1321 gcctccctgg acatgcagct ggagcaggcc cagggaaccc gggagcggct ggcccaagcc 1381 gagtgtgtgc tggaacagtt cggcaatgag gaccaccaca agttcatccg gaagttccac 1441 tccatggcct ccagagcaga gatgccgcag gcccggccct tagaaggcgc attcagcccc 1501 atctccttca agccaggcct ccaccaggct gacatcaagc tgaccgtgtg gaaaaggctc 1561 ttccggaaag ttttgccagc cccggagcct ctcaagttgg accctgccac tgcccaccca 1621 ctcctggagc tctccaaggg caacacggtg gtgcagtgcg ggcttctggc ccagcggcga 1681 gccagccagc ctgagcgctt cgactacagc acctgcgtcc tggccagccg cggcttctcc 1741 tgcggccgcc actactggga ggtggtggtg ggcagcaaga gcgactggcg cctgggggtc 1801 atcaagggca cagccagccg taagggcaag ctgaacaggt cccccgagca cggcgtgtgg 1861 ctgatcggcc tgaaggaggg ccgggtgtac gaagcctttg cctgcccccg ggtacccctg 1921 cccgtggccg gccaccccca ccgcatcggg ctctacctgc actatgagca gggcgaactc 1981 accttcttcg atgccgaccg ccccgatgac ctgcggccgc tctacacctt ccaggccgac 2041 ttccagggca agctctaccc catcctggac acctgctggc acgagagggg cagcaactcg 2101 ctgcccatgg tgctgccccc gcccagcggg cctggccccc tcagccccga gcagcccacc 2161 aagctgctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt 2221 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc 2281 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 2341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 2401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 2461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 2521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 2581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 2641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 2701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 2761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 2821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa 2881 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac 2941 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg 3001 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 3061 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 3121 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg 3181 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 3241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 3301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt 3361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 3421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 3481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 3541 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac 3601 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 3661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 3721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 3781 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 3841 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 3901 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 3961 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc 4021 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 4201 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 4501 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 4681 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 4801 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga 4861 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 4921 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 4981 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5041 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5101 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5161 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5221 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5281 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5401 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 5761 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6001 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 6121 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6181 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6241 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6301 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6361 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6421 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6481 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6541 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6601 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6661 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6721 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6781 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6841 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6901 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6961 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7021 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7081 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7141 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7201 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7261 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7321 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc 7381 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta 7441 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa 7501 tctgcttagg //