LOCUS V025143 5525 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025143 VERSION V025143 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 27..330 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 331..534 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" intron 672..768 /note="modified SV40 intron with splice donor and acceptor sites" /label="SV40 intron" CDS 859..1860 /label="Ctsl(NM_009984)" /note="Ctsl(NM_009984)" /gene="Ctsl" CDS 1897..2613 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 2623..2649 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" polyA_signal 2664..2798 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(2909..2925) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 2933..2949 /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(2957..2987) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3002..3023 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(3311..3899) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4070..4930) /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(4931..5035) /gene="bla" /label="AmpR promoter" primer_bind 5509..5525 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc 61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa 241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac 301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac 601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag 661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg 721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa 781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaggcc cgaattcggt 841 cgaccgagat ctgccaccat gaatctttta ctccttttgg ctgtcctctg cttgggaaca 901 gccttagcta ctccaaaatt tgatcaaacc tttagtgcag agtggcacca gtggaagtcc 961 acacacagaa gactgtatgg cacgaatgag gaagagtgga ggagagcgat atgggagaag 1021 aacatgagaa tgatccagct acacaacggg gaatacagca acgggcagca cggcttttcc 1081 atggagatga acgccttcgg tgacatgacc aatgaggaat tcaggcaggt ggtgaatggc 1141 taccgccacc agaagcacaa gaaggggagg ctttttcagg aaccgctgat gcttaagatc 1201 cccaagtctg tggactggag agaaaagggt tgtgtgactc ctgtgaagaa ccagggccag 1261 tgcgggtctt gttgggcgtt tagcgcatcg ggttgcctag aaggacagat gttccttaag 1321 accggcaaac tgatctcact gagtgaacag aaccttgtgg actgttctca cgctcaaggc 1381 aatcagggct gtaacggagg cctgatggat tttgctttcc agtacattaa ggaaaatgga 1441 ggtctggact cggaggagtc ttacccctat gaagcgaagg acggatcttg taaatacaga 1501 gccgagttcg ctgtggctaa tgacacaggg ttcgtggata tccctcagca agagaaagcc 1561 ctcatgaagg ctgtggcgac tgtggggcct atttctgttg ctatggacgc aagccatccg 1621 tctctccagt tctatagttc aggcatctac tatgaaccca actgtagcag caagaacctc 1681 gaccatgggg ttctgttggt gggctatggc tatgaaggaa cagattcaaa taagaataaa 1741 tattggcttg tcaagaacag ctggggaagt gaatggggta tggaaggcta catcaaaata 1801 gccaaagacc gggacaacca ctgtggactt gccaccgcgg ccagctatcc tgtcgtgaat 1861 ctcgagggtg gcggtggatc aggaggaggt ggctctatgg tgagcaaggg cgaggagctg 1921 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 1981 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 2041 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 2101 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 2161 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 2221 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 2281 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 2341 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 2401 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 2461 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 2521 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 2581 gggatcactc tcggcatgga cgagctgtac aagggtaccg gctacccata cgatgttcca 2641 gattacgctt aagcggccgc ggggatccag acatgataag atacattgat gagtttggac 2701 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg 2761 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt 2821 ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgatct 2881 gcaggcatgc tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 2941 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 3001 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 3061 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 3121 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 3181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 3241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 3301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 3361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 3421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 3481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 3541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 3601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 3661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 3721 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 3781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 3841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 3901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 3961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 4021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 4081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 4141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 4201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 4261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 4321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 4381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 4441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 4501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 4561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 4621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 4681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 4741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 4801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 4861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 4921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 4981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 5041 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 5101 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 5161 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 5221 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 5281 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 5341 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg 5401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 5461 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 5521 ccagt //