LOCUS V026478 6226 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026478 VERSION V026478 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" regulatory 913..922 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 922..987 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 991..1746 /label="Tnfsf15(NM_177371)" /note="Tnfsf15(NM_177371)" /gene="Tnfsf15" polyA_signal 1826..2050 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2096..2524 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2538..2867 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2934..3728 /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 3902..4023 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4072..4088) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4096..4112 /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(4120..4150) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4165..4186 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4474..5059) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5230..6090) /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(6091..6195) /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac 961 atcgactaca aggatgacga tgacaagctt atggcagagg agctggggtt gggcttcgga 1021 gaaggagtcc cagtggaagt gctgccggaa ggctgtagac acaggccaga ggccagggcc 1081 gggctagctg ccaggagcaa agcctgcctg gctctcacct gctgcctgtt gtcatttccc 1141 atcctcgcag gacttagcac cctcctaatg gctggccagc tccgggtccc cggaaaagac 1201 tgtatgcttc gggccataac agaagagaga tctgagcctt caccacagca agtttactca 1261 cctcccagag gcaagccgag agcacacctg acaattaaga aacaaacccc agcaccacat 1321 ctgaaaaatc agctctctgc tctacactgg gaacatgacc tagggatggc cttcaccaag 1381 aacgggatga agtacatcaa caaatccctg gtgatcccag agtcaggaga ctatttcatc 1441 tactcccaga tcacattccg agggaccaca tctgtgtgtg gtgacatcag tcgggggaga 1501 cgaccaaaca agccagactc catcaccatg gttatcacca aggtagcaga cagctaccct 1561 gagcctgccc gcctactaac agggtccaag tctgtgtgtg aaataagcaa caactggttc 1621 cagtccctct accttggggc cacgttctcc ttggaagaag gagacagact aatggtaaac 1681 gtcagtgaca tctccttggt ggattacaca aaagaagata aaactttctt tggagctttc 1741 ttgctataag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc 1801 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 1861 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 1921 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 1981 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 2041 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 2101 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 2161 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 2221 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 2281 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 2341 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 2401 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 2461 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 2521 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 2881 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 2941 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3001 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3061 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3121 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3181 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3241 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3301 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3361 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3421 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3481 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3541 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3601 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3661 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 3721 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 3781 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 3841 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 3901 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3961 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4021 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 4141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 4201 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4261 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4981 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg 5041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 5521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 5581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 5641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 5701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 5761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 5821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 5881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 5941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //