LOCUS V025084 7268 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025084 VERSION V025084 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 4..383 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 386..661 /label="chicken β-actin promoter" intron 662..1674 /note="chimera between introns from chicken β-actin and rabbit β-globin" /label="chimeric intron" regulatory 1681..1690 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 1690..1755 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 1762..3477 /label="ADRA1D(NM_000678)" /note="ADRA1D(NM_000678)" /gene="ADRA1D" CDS 3484..3504 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 3505..4218 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" /label="EGFP" CDS 4219..4242 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 4261..4284 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" polyA_signal 4356..4411 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4771..4787) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4795..4811 /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(4819..4849) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4864..4885 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4944..5139 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 5145..5279 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(5518..6106) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6277..7137) /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(7138..7242) /gene="bla" /label="AmpR promoter" ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc 1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag 1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc 1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc 1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg 1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 1621 ggcggctcta gtgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1681 gccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag 1741 gatgacgatg acaagctcga gatgactttc cgcgatctcc tgagcgtcag tttcgaggga 1801 ccccgcccgg acagcagcgc agggggctcc agcgcgggcg gcggcggggg cagcgcgggc 1861 ggcgcggccc cctcggaggg cccggcggtg ggcggcgtgc cggggggcgc gggcggcggc 1921 ggcggcgtgg tgggcgcagg cagcggcgag gacaaccgga gctccgcggg ggagccgggg 1981 agcgcgggcg cgggcggcga cgtgaatggc acggcggccg tcgggggact ggtggtgagc 2041 gcgcagggcg tgggcgtggg cgtcttcctg gcagccttca tccttatggc cgtggcaggt 2101 aacctgcttg tcatcctctc agtggcctgc aaccgccacc tgcagaccgt caccaactat 2161 ttcatcgtga acctggccgt ggccgacctg ctgctgagcg ccaccgtact gcccttctcg 2221 gccaccatgg aggttctggg cttctgggcc tttggccgcg ccttctgcga cgtatgggcc 2281 gccgtggacg tgctgtgctg cacggcctcc atcctcagcc tctgcaccat ctccgtggac 2341 cggtacgtgg gcgtgcgcca ctcactcaag tacccagcca tcatgaccga gcgcaaggcg 2401 gccgccatcc tggccctgct ctgggtcgta gccctggtgg tgtccgtagg gcccctgctg 2461 ggctggaagg agcccgtgcc ccctgacgag cgcttctgcg gtatcaccga ggaggcgggc 2521 tacgctgtct tctcctccgt gtgctccttc tacctgccca tggcggtcat cgtggtcatg 2581 tactgccgcg tgtacgtggt cgcgcgcagc accacgcgca gcctcgaggc gggcgtcaag 2641 cgcgagcgag gcaaggcctc cgaggtggtg ctgcgcatcc actgtcgcgg cgcggccacg 2701 ggcgccgacg gggcgcacgg catgcgcagc gccaagggcc acaccttccg cagctcgctc 2761 tccgtgcgcc tgctcaagtt ctcccgtgag aagaaagcgg ccaagactct ggccatcgtc 2821 gtgggtgtct tcgtgctctg ctggttccct ttcttctttg tcctgccgct cggctccttg 2881 ttcccgcagc tgaagccatc ggagggcgtc ttcaaggtca tcttctggct cggctacttc 2941 aacagctgcg tgaacccgct catctacccc tgttccagcc gcgagttcaa gcgcgccttc 3001 ctccgtctcc tgcgctgcca gtgccgtcgt cgccggcgcc gccgccctct ctggcgtgtc 3061 tacggccacc actggcgggc ctccaccagc ggcctgcgcc aggactgcgc cccgagttcg 3121 ggcgacgcgc cccccggagc gccgctggcc ctcaccgcgc tccccgaccc cgaccccgaa 3181 cccccaggca cgcccgagat gcaggctccg gtcgccagcc gtcgaaagcc acccagcgcc 3241 ttccgcgagt ggaggctgct ggggccgttc cggagaccca cgacccagct gcgcgccaaa 3301 gtctccagcc tgtcgcacaa gatccgcgcc gggggcgcgc agcgcgcaga ggcagcgtgc 3361 gcccagcgct cagaggtgga ggctgtgtcc ctaggcgtcc cacacgaggt ggccgagggc 3421 gccacctgcc aggcctacga attggccgac tacagcaacc tacgggagac cgatattgaa 3481 ttcgaaaacc tgtacttcca atccgtgagc aagggcgagg agctgttcac cggggtggtg 3541 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 3601 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 3661 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 3721 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 3781 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 3841 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 3901 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 3961 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 4021 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 4081 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 4141 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 4201 atggacgagc tgtacaagtg gagccacccg cagttcgaga agggcggcgg cagcagcgcc 4261 tggagccacc cgcagttcga gaagtgaaga tctttttccc tctgccaaaa attatgggga 4321 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc 4381 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca 4441 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc 4501 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 4561 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 4621 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 4681 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc 4741 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4801 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4861 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4921 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta 4981 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 5041 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 5101 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag 5161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc 5281 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 5341 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 5401 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 5461 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 5521 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 5581 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5641 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5701 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5761 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 5821 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 5881 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 5941 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6001 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 6061 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6121 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6181 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6241 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6301 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6361 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6421 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6481 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6541 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6601 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6661 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6721 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6781 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6841 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6901 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6961 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7021 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7081 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7141 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7201 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7261 gccacctg //