LOCUS V020214 7214 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020214 VERSION V020214 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..1894 /label="UBA5(NM_024818)" /note="UBA5(NM_024818)" /gene="UBA5" CDS 1901..1966 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1990..2578 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2644..2692 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2894..3322 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3336..3665 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3732..4526 /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 4702..4823 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4872..4888) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4896..4912 /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(4920..4950) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4965..4986 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5274..5862) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6033..6893) /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(6894..6998) /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 ccatggcgga gtctgtggag cgcctgcagc agcgggtcca 721 ggagctggag cgggaacttg cccaggagag gagtctgcag gtcccgagga gcggcgacgg 781 agggggcggc cgggtccgca tcgagaagat gagctcagag gtggtggatt cgaatcccta 841 cagccgcttg atggcattga aacgaatggg aattgtaagc gactatgaga aaatccgtac 901 ctttgccgta gcaatagtag gtgttggtgg agtaggtagt gtgactgctg aaatgctgac 961 aagatgtggc attggtaagt tgctactctt tgattatgac aaggtggaac tagccaatat 1021 gaatagactt ttcttccaac ctcatcaagc aggattaagt aaagttcaag cagcagaaca 1081 tactctgagg aacattaatc ctgatgttct ttttgaagta cacaactata atataaccac 1141 agtggaaaac tttcaacatt tcatggatag aataagtaat ggtgggttag aagaaggaaa 1201 acctgttgat ctagttctta gctgtgtgga caattttgaa gctcgaatga caataaatac 1261 agcttgtaat gaacttggac aaacatggat ggaatctggg gtcagtgaaa atgcagtttc 1321 agggcatata cagcttataa ttcctggaga atctgcttgt tttgcgtgtg ctccaccact 1381 tgtagttgct gcaaatattg atgaaaagac tctgaaacga gaaggtgttt gtgcagccag 1441 tcttcctacc actatgggtg tggttgctgg gatcttagta caaaacgtgt taaagtttct 1501 gttaaatttt ggtactgtta gtttttacct tggatacaat gcaatgcagg atttttttcc 1561 tactatgtcc atgaagccaa atcctcagtg tgatgacaga aattgcagga agcagcagga 1621 ggaatataag aaaaaggtag cagcactgcc taaacaagag gttatacaag aagaggaaga 1681 gataatccat gaagataatg aatggggtat tgagctggta tctgaggttt cagaagagga 1741 actgaaaaat ttttcaggtc cagttccaga cttacctgaa ggaattacag tggcatacac 1801 aattccaaaa aagcaagaag attctgtcac tgagttaaca gtggaagatt ctggtgaaag 1861 cttggaagac ctcatggcca aaatgaagaa tatgctcgag gactacaaag accatgacgg 1921 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 1981 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 2041 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 2101 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 2161 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 2221 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 2281 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 2341 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 2401 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 2461 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 2521 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 2581 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 2641 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 2701 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 2761 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 2821 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 2881 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2941 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 3061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3121 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3241 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 3301 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3481 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3601 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3661 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3721 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3781 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3841 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3901 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3961 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4021 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4081 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4141 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4201 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4261 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4321 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4381 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4441 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4501 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 4561 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 4621 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4681 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 4741 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4801 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 4861 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 5041 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 5101 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 5161 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 5221 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5281 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 5341 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 5401 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5461 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5521 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5581 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 5641 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5701 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5761 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5821 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 5881 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 5941 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6001 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6061 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6121 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6181 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6241 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6301 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6361 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6421 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6481 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6541 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6601 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6661 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6721 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6781 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6841 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 6901 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 6961 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7021 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 7081 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 7141 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 7201 agaatctgct tagg //