LOCUS V018573 7028 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018573 VERSION V018573 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..1675 /label="NUDC(NM_006600)" /note="NUDC(NM_006600)" /gene="NUDC" CDS 1715..1780 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1804..2392 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2458..2506 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2708..3136 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3150..3479 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3546..4340 /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 4516..4637 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4686..4702) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4710..4726 /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(4734..4764) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4779..4800 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5088..5676) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5847..6707) /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(6708..6812) /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 ccatgggcgg agagcaggag gaggagcggt tcgacggcat 721 gttgctggcc atggctcagc agcacgaggg cggcgtgcag gagcttgtga acaccttctt 781 cagcttcctt cgacgcaaaa cagacttttt cattggagga gaagaaggga tggcagagaa 841 gcttatcaca cagactttca gccaccacaa tcagctggca cagaagaccc ggcgggagaa 901 gagagcccgg caggaggccg agcggcggga gaaggcggag cgggcggcca gactggccaa 961 ggaagccaag tcagagacct cagggcccca gatcaaggag ctaactgatg aagaggcaga 1021 gaggctgcag ctagagattg accagaaaaa ggatgcagag aatcatgagg cccagctcaa 1081 gaacggcagc cttgactccc cagggaagca ggatactgag gaagatgagg aggaagatga 1141 gaaggacaaa ggaaaactga agcccaacct aggcaacggg gcagacctgc ccaattaccg 1201 ctggacccag accctgtcgg agctggacct ggcggtccct ttctgtgtga acttccggct 1261 gaaagggaag gacatggtgg tggacatcca gcggcggcac ctccgggtgg ggctcaaggg 1321 gcagccagcg atcattgatg gggagctcta caatgaagtg aaggtggagg agagctcgtg 1381 gctcattgag gacggcaagg tggtgactgt gcatctggag aagatcaata agatggagtg 1441 gtggagccgc ttggtgtcca gtgaccctga gatcaacacc aagaagatta accctgagaa 1501 ttccaagctg tcagacctgg acagtgagac tcgcagcatg gtggaaaaga tgatgtatga 1561 ccagcgacag aagtccatgg ggctgccaac ttcagacgaa cagaagaaac aggagattct 1621 gaagaagttc atggatcaac atccggagat ggatttttcc aaggctaaat tcaacgaatt 1681 ctgcagatat ccagcacagt ggcggccgct cgaggactac aaagaccatg acggtgatta 1741 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 1801 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 1861 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 1921 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 1981 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 2041 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 2101 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 2161 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 2221 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 2281 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 2341 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 2401 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 2461 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 2521 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2581 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2641 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 2701 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2761 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2821 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 2881 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 2941 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 3001 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 3061 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 3121 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3301 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3361 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3421 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3481 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3541 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3601 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3661 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3721 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3781 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3841 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3901 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 3961 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 4021 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 4081 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 4141 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4201 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4261 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 4321 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 4381 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4441 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4501 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4561 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4621 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4921 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5281 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5521 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5881 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5941 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 6001 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 6061 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 6121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6721 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6781 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6841 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 6901 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 6961 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 7021 tgcttagg //