LOCUS V019674 7631 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019674 VERSION V019674 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..2311 /label="EGR1(NM_001964)" /note="EGR1(NM_001964)" /gene="EGR1" CDS 2318..2383 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2407..2995 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3061..3109 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3311..3739 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3753..4082 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4149..4943 /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 5119..5240 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5289..5305) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5313..5329 /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(5337..5367) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5382..5403 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5691..6279) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6450..7310) /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(7311..7415) /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 ccatggccgc ggccaaggcc gagatgcagc tgatgtcccc 721 gctgcagatc tctgacccgt tcggatcctt tcctcactcg cccaccatgg acaactaccc 781 taagctggag gagatgatgc tgctgagcaa cggggctccc cagttcctcg gcgccgccgg 841 ggccccagag ggcagcggca gcaacagcag cagcagcagc agcgggggcg gtggaggcgg 901 cgggggcggc agcaacagca gcagcagcag cagcaccttc aaccctcagg cggacacggg 961 cgagcagccc tacgagcacc tgaccgcaga gtcttttcct gacatctctc tgaacaacga 1021 gaaggtgctg gtggagacca gttaccccag ccaaaccact cgactgcccc ccatcaccta 1081 tactggccgc ttttccctgg agcctgcacc caacagtggc aacaccttgt ggcccgagcc 1141 cctcttcagc ttggtcagtg gcctagtgag catgaccaac ccaccggcct cctcgtcctc 1201 agcaccatct ccagcggcct cctccgcctc cgcctcccag agcccacccc tgagctgcgc 1261 agtgccatcc aacgacagca gtcccattta ctcagcggca cccaccttcc ccacgccgaa 1321 cactgacatt ttccctgagc cacaaagcca ggccttcccg ggctcggcag ggacagcgct 1381 ccagtacccg cctcctgcct accctgccgc caagggtggc ttccaggttc ccatgatccc 1441 cgactacctg tttccacagc agcaggggga tctgggcctg ggcaccccag accagaagcc 1501 cttccagggc ctggagagcc gcacccagca gccttcgcta acccctctgt ctactattaa 1561 ggcctttgcc actcagtcgg gctcccagga cctgaaggcc ctcaatacca gctaccagtc 1621 ccagctcatc aaacccagcc gcatgcgcaa gtaccccaac cggcccagca agacgccccc 1681 ccacgaacgc ccttacgctt gcccagtgga gtcctgtgat cgccgcttct cccgctccga 1741 cgagctcacc cgccacatcc gcatccacac aggccagaag cccttccagt gccgcatctg 1801 catgcgcaac ttcagccgca gcgaccacct caccacccac atccgcaccc acacaggcga 1861 aaagcccttc gcctgcgaca tctgtggaag aaagtttgcc aggagcgatg aacgcaagag 1921 gcataccaag atccacttgc ggcagaagga caagaaagca gacaaaagtg ttgtggcctc 1981 ttcggccacc tcctctctct cttcctaccc gtccccggtt gctacctctt acccgtcccc 2041 ggttactacc tcttatccat ccccggccac cacctcatac ccatcccctg tgcccacctc 2101 cttctcctct cccggctcct cgacctaccc atcccctgtg cacagtggct tcccctcccc 2161 gtcggtggcc accacgtact cctctgttcc ccctgctttc ccggcccagg tcagcagctt 2221 cccttcctca gctgtcacca actccttcag cgcctccaca gggctttcgg acatgacagc 2281 aaccttttct cccaggacaa ttgaaatttg cctcgaggac tacaaagacc atgacggtga 2341 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat 2401 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 2461 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 2521 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 2581 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 2641 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 2701 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 2761 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 2821 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 2881 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 2941 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 3001 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 3061 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg 3121 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac 3181 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc 3241 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg 3301 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 3361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 3421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg 3481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 3541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 3601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 3661 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa 3721 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 4021 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 4081 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 4141 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 4201 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 4261 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 4321 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 4381 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 4441 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 4501 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 4561 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 4621 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 4681 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4741 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4801 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4861 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4921 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg 4981 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 5041 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 5101 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat 5161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 5221 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg 5281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 5341 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 5401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 5461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 5521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 5581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 5641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 5701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 5761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 5821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 5881 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 5941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 6001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 6061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 6121 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 6181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 6241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 6301 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 6361 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 6421 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 6481 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 6541 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 6601 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 6661 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 6721 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 6781 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 6841 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 6901 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 6961 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 7021 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 7081 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 7141 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 7201 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 7261 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 7321 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 7381 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 7441 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct 7501 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt 7561 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga 7621 atctgcttag g //