LOCUS V019615 7304 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019615 VERSION V019615 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..1984 /label="ENO2(NM_001975)" /note="ENO2(NM_001975)" /gene="ENO2" CDS 1991..2056 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2080..2668 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2734..2782 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2984..3412 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3426..3755 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3822..4616 /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 4792..4913 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4962..4978) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4986..5002 /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(5010..5040) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5055..5076 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5364..5952) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6123..6983) /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(6984..7088) /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 ccatgtccat agagaagatc tgggcccggg agatcctgga 721 ctcccgcggg aaccccacag tggaggtgga tctctatact gccaaaggtc ttttccgggc 781 tgcagtgccc agtggagcct ctacgggcat ctatgaggcc ctggagctga gggatggaga 841 caaacagcgt tacttaggca aaggtgtcct gaaggcagtg gaccacatca actccaccat 901 cgcgccagcc ctcatcagct caggtctctc tgtggtggag caagagaaac tggacaacct 961 gatgctggag ttggatggga ctgagaacaa atccaagttt ggggccaatg ccatcctggg 1021 tgtgtctctg gccgtgtgta aggcaggggc agctgagcgg gaactgcccc tgtatcgcca 1081 cattgctcag ctggccggga actcagacct catcctgcct gtgccggcct tcaacgtgat 1141 caatggtggc tctcatgctg gcaacaagct ggccatgcag gagttcatga tcctcccagt 1201 gggagctgag agctttcggg atgccatgcg actaggtgca gaggtctacc atacactcaa 1261 gggagtcatc aaggacaaat acggcaagga tgccaccaat gtgggggatg aaggtggctt 1321 tgcccccaat atcctggaga acagtgaagc cttggagctg gtgaaggaag ccatcgacaa 1381 ggctggctac acggaaaaga tcgttattgg catggatgtt gctgcctcag agttttatcg 1441 tgatggcaaa tatgacttgg acttcaagtc tcccactgat ccttcccgat acatcactgg 1501 ggaccagctg ggggcactct accaggactt tgtcagggac tatcctgtgg tctccattga 1561 ggacccattt gaccaggatg attgggctgc ctggtccaag ttcacagcca atgtagggat 1621 ccagattgtg ggtgatgacc tgacagtgac caacccaaaa cgtattgagc gggcagtgga 1681 agaaaaggcc tgcaactgtc tgctgctcaa ggtcaaccag atcggctcgg tcactgaagc 1741 catccaagcg tgcaagctgg cccaggagaa tggctggggg gtcatggtga gtcatcgctc 1801 aggagagact gaggacacat tcattgctga cctggtggtg gggctgtgca caggccagat 1861 caagactggt gccccgtgcc gttctgaacg tctggctaaa tacaaccagc tcatgagaat 1921 tgaggaagag ctgggggatg aagctcgctt tgccggacat aacttccgta atcccagtgt 1981 gctgctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 2041 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 2101 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 2161 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 2221 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 2281 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 2341 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 2401 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 2461 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 2521 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 2581 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 2641 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 2701 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 2761 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 2821 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 2881 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 2941 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 3001 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3061 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3121 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 3181 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3241 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3301 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 3361 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3661 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3721 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3781 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3841 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3901 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3961 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 4021 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4081 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4141 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4201 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4261 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4321 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4381 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4441 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4501 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4561 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4621 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 4681 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 4741 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 4801 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4861 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4921 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 4981 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5041 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5101 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5161 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5221 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5281 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5341 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5401 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5461 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5521 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 5581 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5641 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5701 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5761 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 5821 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5881 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5941 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6001 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6061 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6121 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6181 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6241 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6301 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6361 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6421 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6481 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6541 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6601 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6661 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6721 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6781 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6841 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6901 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6961 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7021 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7081 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 7141 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 7201 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 7261 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //