LOCUS V024688 6646 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024688 VERSION V024688 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..2166 /label="LUC7L2(NM_016019)" /note="LUC7L2(NM_016019)" /gene="LUC7L2" polyA_signal 2246..2470 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2516..2944 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2958..3287 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3354..4148 /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 4322..4443 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4492..4508) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4516..4532 /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(4540..4570) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4585..4606 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4894..5479) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5650..6510) /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(6511..6615) /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 atgtcggcgc aggcccagat gcgcgcgatg 1021 ctggaccagt tgatgggcac ctcccgggac ggagatacaa ctcgtcaacg aatcaaattc 1081 agtgatgaca gagtatgcaa gagtcacctt ctcaactgtt gtcctcatga tgtcctttct 1141 ggaactagaa tggatcttgg agaatgtctg aaagtccatg acctggcttt aagagcggat 1201 tatgaaattg catccaaaga acaagatttt ttctttgaac ttgatgccat ggatcatctg 1261 cagtcattca ttgcagattg tgatcgtaga acagaagtgg ccaagaaaag attagcagaa 1321 actcaagaag agattagtgc tgaagtagca gcaaaggcag aacgtgttca tgagttaaat 1381 gaagaaattg gtaaattgtt agccaaggtg gaacaactag gagctgaagg gaatgtggag 1441 gaatcccaga aagtaatgga tgaagtagag aaagcacggg caaagaaaag agaagcagag 1501 gaagtttatc ggaattctat gccagcttcc agttttcagc agcagaaact tcgagtctgt 1561 gaagtctgct ctgcctattt aggacttcat gataatgaca gacgactggc tgatcatttt 1621 gggggtaaac tgcacctggg atttattgaa ataagagaga agcttgaaga attaaagaga 1681 gtcgtagctg agaagcagga gaaaagaaac caggaacggc tgaaacgaag agaagagaga 1741 gagagagaag aaagggagaa gctgaggagg tcccgatcac acagcaagaa tccaaaaaga 1801 tccaggtcca gagagcatcg cagacatcga tctcgctcca tgtcacgtga acgcaagagg 1861 agaactcgat ccaaatctcg ggagaaacgc catcgccaca ggtcccgctc cagcagccgt 1921 agccgcagcc gtagccacca gagaagtcgg cacagttcta gagataggag cagagaacga 1981 tccaagagga gatcctcaaa agaaagattc agagaccaag acttagcatc atgtgacaga 2041 gacaggagtt caagagacag atcacctcgt gacagagatc ggaaagataa gaagcggtcc 2101 tatgagagtg ctaatggcag atcagaagac aggaggagct ctgaagagcg cgaagcaggg 2161 gagatctaag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc 2221 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 2281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 2341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 2401 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 2461 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 2881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 2941 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3301 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3361 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3421 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3481 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3541 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3601 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3661 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3721 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3781 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3841 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3901 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3961 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4021 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4081 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4141 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 4201 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4261 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4321 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4381 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4441 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4501 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 4561 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 4621 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4681 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4741 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4801 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4861 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4921 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4981 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5041 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 5101 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5161 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 5221 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5281 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5341 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5401 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg 5461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5701 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 5941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 6001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 6061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 6301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 6361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //