LOCUS V031952 6266 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031952 VERSION V031952 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 941..1756 /label="CAPZB(NM_004930)" /note="CAPZB(NM_004930)" /gene="CAPZB" CDS 1763..1828 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1866..2090 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2136..2564 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2578..2907 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2974..3768 /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 3942..4063 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4112..4128) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4136..4152 /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(4160..4190) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4205..4226 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4514..5099) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5270..6130) /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(6131..6235) /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 atccgccacc atgagtgatc agcagctgga 961 ctgtgccttg gacctaatga ggcgcctgcc tccccagcaa atcgagaaaa acctcagcga 1021 cctgatcgac ctggtcccca gtctatgtga ggatctcctg tcttctgttg accagccact 1081 gaaaattgcc agagacaagg tggtgggaaa ggattacctt ttgtgtgact acaacagaga 1141 tggggactcc tataggtcac catggagtaa caagtatgac cctcccttgg aggatggggc 1201 catgccgtca gctcggctga gaaagctgga ggtggaagcc aacaatgcct ttgaccagta 1261 tcgagacctg tattttgaag gtggcgtctc atctgtctac ctctgggatc tggatcatgg 1321 ctttgctgga gtgatcctca taaagaaggc tggagatgga tcaaagaaga tcaaaggctg 1381 ctgggattcc atccacgtgg tagaagtgca ggagaaatcc agcggtcgca ccgcccatta 1441 caagttgacc tccacggtga tgctgtggct gcagaccaac aaatctggct ctggcaccat 1501 gaacctcgga ggcagcctta ccagacagat ggagaaggat gaaactgtga gtgactgctc 1561 cccacacata gccaacatcg ggcgcctggt agaggacatg gaaaataaaa tcagaagtac 1621 gctgaacgag atctactttg gaaaaacaaa ggatatcgtc aatgggctga ggtctgtgca 1681 gacttttgca gacaaatcaa aacaagaagc tctgaagaat gacctggtgg aggctttgaa 1741 gagaaagcag caatgcctcg aggactacaa agaccatgac ggtgattata aagatcatga 1801 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 1861 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 1921 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 1981 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 2041 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 2101 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 2161 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 2221 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 2281 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 2341 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 2401 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 2461 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 2521 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 2581 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2641 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 2701 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 2761 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 2821 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 2881 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 2941 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3001 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3061 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3121 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3181 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 3241 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3301 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3361 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 3421 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 3481 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 3541 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 3601 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 3661 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 3721 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 3781 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 3841 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 3901 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 3961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4081 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 4981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5041 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5101 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5161 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5221 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5281 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5341 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5401 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5461 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5521 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5581 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5641 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5701 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 5761 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 5821 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 5881 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 5941 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6001 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6061 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6121 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6181 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6241 ttccccgaaa agtgccacct gacgtc //