LOCUS V025172 6938 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025172 VERSION V025172 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..3147 /label="ADRA2C(NM_000683)" /note="ADRA2C(NM_000683)" /gene="ADRA2C" CDS 3154..3174 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 3175..3888 /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 3889..3912 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3931..3954 /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 4026..4081 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4441..4457) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4465..4481 /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(4489..4519) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4534..4555 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4614..4809 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4815..4949 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(5188..5776) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5947..6807) /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(6808..6912) /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 gatggcgtcc ccggcgctgg cggcggcgct ggcggtggcg 1801 gcagcggcgg gccccaatgc gagcggcgcg ggcgagaggg gcagcggcgg ggttgccaat 1861 gcctcggggg cttcctgggg gccgccgcgc ggccagtact cggcgggcgc ggtggcaggg 1921 ctggctgccg tggtgggctt cctcatcgtc ttcaccgtgg tgggcaacgt gctggtggtg 1981 atcgccgtgc tgaccagccg ggcgctgcgc gcgccacaga acctcttcct ggtgtcgctg 2041 gcctcggccg acatcctggt ggccacgctg gtcatgccct tctcgttggc caacgagctc 2101 atggcctact ggtacttcgg gcaggtgtgg tgcggcgtgt acctggcgct cgatgtgctg 2161 ttttgcacct cgtcgatcgt gcatctgtgt gccatcagcc tggaccgcta ctggtcggtg 2221 acgcaggccg tcgagtacaa cctgaagcgc acaccacgcc gcgtcaaggc caccatcgtg 2281 gccgtgtggc tcatctcggc cgtcatctcc ttcccgccgc tggtctcgct ctaccgccag 2341 cccgacggcg ccgcctaccc gcagtgcggc ctcaacgacg agacctggta catcctgtcc 2401 tcctgcatcg gctccttctt cgcgccctgc ctcatcatgg gcctggtcta cgcgcgcatc 2461 taccgagtgg ccaagctgcg cacgcgcacg ctcagcgaga agcgcgcccc cgtgggcccc 2521 gacggtgcgt ccccgactac cgaaaacggg ctgggcgcgg cggcaggcgc aggcgagaac 2581 gggcactgcg cgcccccgcc cgccgacgtg gagccggacg agagcagcgc agcggccgag 2641 aggcggcggc gccggggcgc gttgcggcgg ggcgggcggc ggcgagcggg cgcggagggg 2701 ggcgcgggcg gtgcggacgg gcagggggcg gggccggggg cggctgagtc gggggcgctg 2761 accgcctcca ggtccccggg gcccggtggc cgcctgtcgc gcgccagctc gcgctccgtc 2821 gagttcttcc tgtcgcgccg gcgccgggcg cgcagcagcg tgtgccgccg caaggtggcc 2881 caggcgcgcg agaagcgctt cacctttgtg ctggctgtgg tcatgggcgt gttcgtgctc 2941 tgctggttcc ccttcttctt cagctacagc ctgtacggca tctgccgcga ggcctgccag 3001 gtgcccggcc cgctcttcaa gttcttcttc tggatcggct actgcaacag ctcgctcaac 3061 ccggtcatct acacggtctt caaccaggat ttccggcgat cctttaagca catcctcttc 3121 cgacggagga gaaggggctt caggcaggaa ttcgaaaacc tgtacttcca atccgtgagc 3181 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta 3241 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg 3301 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc 3361 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac 3421 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac 3481 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc 3541 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag 3601 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag 3661 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac 3721 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc 3781 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag 3841 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg gagccacccg 3901 cagttcgaga agggcggcgg cagcagcgcc tggagccacc cgcagttcga gaagtgaaga 3961 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc 4021 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc 4081 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta 4141 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggtggct ataaagaggt 4201 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact 4261 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa 4321 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata 4381 gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg 4441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc 4621 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 4681 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 4741 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 4801 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4861 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4921 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg 4981 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 5041 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 5101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 5161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 5281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 5401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 5521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 5581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 5641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 5701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 5761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5941 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 6001 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 6061 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6121 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6181 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6241 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6301 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6361 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6421 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6481 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6541 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6601 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6661 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6721 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6781 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6901 aggggttccg cgcacatttc cccgaaaagt gccacctg //