LOCUS V018363 7283 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018363 VERSION V018363 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..1963 /label="DDB2(NM_000107)" /note="DDB2(NM_000107)" /gene="DDB2" CDS 1970..2035 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2059..2647 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2713..2761 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2963..3391 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3405..3734 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3801..4595 /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 4771..4892 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4941..4957) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4965..4981 /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(4989..5019) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5034..5055 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5343..5931) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6102..6962) /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(6963..7067) /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 ccatggctcc caagaaacgc ccagaaaccc agaagacctc 721 cgagattgta ttacgcccca ggaacaagag gagcaggagt cccctggagc tggagcccga 781 ggccaagaag ctctgtgcga agggctccgg tcctagcaga agatgtgact cagactgcct 841 ctgggtgggg ctggctggcc cacagatcct gccaccatgc cgcagcatcg tcaggaccct 901 ccaccagcat aagctgggca gagcttcctg gccatctgtc cagcaggggc tccagcagtc 961 ctttttgcac actctggatt cttaccggat attacaaaag gctgccccct ttgacaggag 1021 ggctacatcc ttggcgtggc acccaactca ccccagcacc gtggctgtgg gttccaaagg 1081 gggagatatc atgctctgga attttggcat caaggacaaa cccaccttca tcaaagggat 1141 tggagctgga gggagcatca ctgggctgaa gtttaaccct ctcaatacca accagtttta 1201 cgcctcctca atggagggaa caactaggct gcaagacttt aaaggcaaca ttctacgagt 1261 ttttgccagc tcagacacca tcaacatctg gttttgtagc ctggatgtgt ctgctagtag 1321 ccgaatggtg gtcacaggag acaacgtggg gaacgtgatc ctgctgaaca tggacggcaa 1381 agagctttgg aatctcagaa tgcacaaaaa gaaagtgacg catgtggccc tgaacccatg 1441 ctgtgattgg ttcctggcca cagcctccgt agatcaaaca gtgaaaattt gggacctgcg 1501 ccaggttaga gggaaagcca gcttcctcta ctcgctgccg cacaggcatc ctgtcaacgc 1561 agcttgtttc agtcccgatg gagcccggct cctgaccacg gaccagaaga gcgagatccg 1621 agtttactct gcttcccagt gggactgccc cctgggcctg atcccgcacc ctcaccgtca 1681 cttccagcac ctcacaccca tcaaggcagc ctggcatcct cgctacaacc tcattgttgt 1741 gggccgatac ccagatccta atttcaaaag ttgtacccct tatgaattga ggacgatcga 1801 cgtgttcgat ggaaactcag ggaagatgat gtgtcagctc tatgacccag aatcttctgg 1861 catcagttcg cttaatgaat tcaatcccat gggggacacg ctggcctctg caatgggtta 1921 ccacattctc atctggagcc aggaggaagc caggacacgg aagctcgagg actacaaaga 1981 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg 2041 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 2101 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 2161 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 2221 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 2281 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 2341 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2401 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 2461 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 2521 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 2581 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 2641 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa 2701 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt 2761 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat 2821 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa 2881 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc 2941 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 3001 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 3061 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc 3121 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 3181 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3241 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3301 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat 3361 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 3421 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3481 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3541 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3601 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3661 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3721 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 3781 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3841 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3901 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3961 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 4021 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 4081 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 4141 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 4201 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 4261 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4321 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4381 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4441 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4501 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4561 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg 4621 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 4681 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4741 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 4801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 4921 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4981 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 5041 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 5101 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 5161 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 5221 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 5281 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 5341 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 5401 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 5461 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 5521 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5581 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 5641 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5701 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5761 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 5821 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5881 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5941 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 6001 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 6061 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 6121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 6181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 6241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 6301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 6361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 6421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 6481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 6661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 6721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 6781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 6841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 7021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 7081 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca 7141 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg 7201 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca 7261 attgcatgaa gaatctgctt agg //