LOCUS V043273 7456 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043273 VERSION V043273 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..2946 /label="F13B(NM_001994)" /note="F13B(NM_001994)" /gene="F13B" CDS 2953..3018 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3056..3280 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3326..3754 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3768..4097 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4164..4958 /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 5132..5253 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5302..5318) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5326..5342 /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(5350..5380) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5395..5416 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5704..6289) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6460..7320) /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(7321..7425) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatgaggt tgaaaaacct gacttttatc atcatattga taatctcagg agaactctat 1021 gcagaagaga aaccctgtgg ttttcctcat gtggaaaatg gaagaattgc ccaatattac 1081 tatactttta aaagctttta ctttccaatg agcatagaca aaaaattgtc atttttctgc 1141 ttggctggtt ataccactga aagtggaaga caagaagagc aaaccacgtg tacaacagaa 1201 ggctggtctc cagagccaag gtgcttcaaa aaatgcacta agcctgacct gagtaatggt 1261 tacatctctg atgtaaagtt attgtataaa attcaagaga acatgcgtta tggttgcgct 1321 tcagggtaca aaaccactgg agggaaggat gaagaagtgg ttcaatgtct ctctgatgga 1381 tggtcttctc aaccaacctg taggaaagaa catgaaacat gtttggctcc tgaattatat 1441 aatggaaatt attccacaac acagaaaaca ttcaaagtga aggacaaagt acaatacgaa 1501 tgtgctactg gctactacac agctggagga aagaagacag aggaggtaga atgtctcaca 1561 tacggatggt ctctcacacc aaaatgtacc aaattaaagt gctcttcttt aagattaatt 1621 gaaaatggtt attttcatcc tgtaaagcaa acctatgaag aaggagatgt cgttcagttt 1681 ttctgtcatg aaaattatta tctaagtgga tctgatttaa ttcaatgcta taactttggt 1741 tggtacccag aatctcctgt atgcgaagga agaagaaaca gatgtcctcc tccacctctg 1801 cccataaact ccaaaattca aacacattca acaacttatc gtcatggaga aatagttcat 1861 atagaatgtg aacttaattt tgagatccat gggtcagcag aaatacgttg tgaagatgga 1921 aaatggacag aacctccaaa atgcattgaa ggacaggaga aggtagcctg tgaggaacca 1981 cccttcattg aaaatggtgc agcaaattta cactctaaga tttattacaa tggggataaa 2041 gtgacatatg catgtaaaag cggctacctt ctccatggat cgaatgagat aacttgtaat 2101 cgtggaaaat ggacacttcc tcctgagtgt gttgaaaata atgagaattg taagcatcct 2161 cctgttgtaa tgaatggggc tgttgcagac gggatattgg caagctatgc aacaggatcc 2221 tcagtggaat atagatgcaa tgaatattac ttactgaggg gatcaaaaat atctcgttgc 2281 gaacaaggaa aatggtcatc cccacctgtt tgcttggaac catgtactgt taatgtggat 2341 tacatgaaca gaaataacat agaaatgaag tggaaatatg aagggaaagt cttacatgga 2401 gatttaatag attttgtatg taaacaggga tatgacttat ctccattaac cccattgtct 2461 gaattatctg tgcagtgcaa cagaggagaa gtgaaatatc ctttatgtac tagaaaagaa 2521 tctaaaggaa tgtgcacatc tcctcctctt attaaacatg gagtcattat tagttcaaca 2581 gtagacacct atgaaaatgg ctcttcagta gaatacagat gttttgatca ccatttccta 2641 gaaggatcta gggaggccta ttgtttagat ggaatgtgga ctacaccacc attgtgttta 2701 gagccatgca cattatcttt tactgaaatg gaaaagaata atttacttct gaaatgggat 2761 tttgacaata gaccacacat tttgcatggt gaatatattg agtttatttg tagaggagat 2821 acttatccag ctgaattata tattactgga tctatactta gaatgcaatg tgacagaggg 2881 cagttaaaat atccaagatg tattccaaga caaagcactc tgtcttatca agaaccctta 2941 agaacactcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac 3001 aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg 3061 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 3121 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 3181 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 3241 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 3301 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 3721 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3841 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3901 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4021 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4081 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 4141 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 4201 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 4261 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 4321 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 4381 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4441 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 4501 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 4561 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 4621 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 4681 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 4741 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 4801 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 4861 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 4921 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 4981 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 5041 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 5101 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 5161 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 5221 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 5281 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7441 agtgccacct gacgtc //