LOCUS V024943 6548 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024943 VERSION V024943 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..2757 /label="ADORA2B(NM_000676)" /note="ADORA2B(NM_000676)" /gene="ADORA2B" CDS 2764..2784 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2785..3498 /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 3499..3522 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3541..3564 /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 3636..3691 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4051..4067) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4075..4091 /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(4099..4129) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4144..4165 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4224..4419 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4425..4559 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(4798..5386) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5557..6417) /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(6418..6522) /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 gatgctgctg gagacacagg acgcgctgta cgtggcgctg 1801 gagctggtca tcgccgcgct ttcggtggcg ggcaacgtgc tggtgtgcgc cgcggtgggc 1861 acggcgaaca ctctgcagac gcccaccaac tacttcctgg tgtccctggc tgcggccgac 1921 gtggccgtgg ggctcttcgc catccccttt gccatcacca tcagcctggg cttctgcact 1981 gacttctacg gctgcctctt cctcgcctgc ttcgtgctgg tgctcacgca gagctccatc 2041 ttcagccttc tggccgtggc agtcgacaga tacctggcca tctgtgtccc gctcaggtat 2101 aaaagtttgg tcacggggac ccgagcaaga ggggtcattg ctgtcctctg ggtccttgcc 2161 tttggcatcg gattgactcc attcctgggg tggaacagta aagacagtgc caccaacaac 2221 tgcacagaac cctgggatgg aaccacgaat gaaagctgct gccttgtgaa gtgtctcttt 2281 gagaatgtgg tccccatgag ctacatggta tatttcaatt tctttgggtg tgttctgccc 2341 ccactgctta taatgctggt gatctacatt aagatcttcc tggtggcctg caggcagctt 2401 cagcgcactg agctgatgga ccactcgagg accaccctcc agcgggagat ccatgcagcc 2461 aagtcactgg ccatgattgt ggggattttt gccctgtgct ggttacctgt gcatgctgtt 2521 aactgtgtca ctcttttcca gccagctcag ggtaaaaata agcccaagtg ggcaatgaat 2581 atggccattc ttctgtcaca tgccaattca gttgtcaatc ccattgtcta tgcttaccgg 2641 aaccgagact tccgctacac ttttcacaaa attatctcca ggtatcttct ctgccaagca 2701 gatgtcaaga gtgggaatgg tcaggctggg gtacagcctg ctctcggtgt gggcctagaa 2761 ttcgaaaacc tgtacttcca atccgtgagc aagggcgagg agctgttcac cggggtggtg 2821 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2881 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2941 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 3001 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 3061 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 3121 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 3181 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 3241 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 3301 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 3361 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 3421 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3481 atggacgagc tgtacaagtg gagccacccg cagttcgaga agggcggcgg cagcagcgcc 3541 tggagccacc cgcagttcga gaagtgaaga tctttttccc tctgccaaaa attatgggga 3601 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc 3661 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca 3721 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc 3781 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 3841 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 3901 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 3961 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc 4021 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4201 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta 4261 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 4321 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 4381 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag 4441 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4501 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc 4561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 4621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 4681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 4741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 4801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 4861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 4921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 4981 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 5101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 5161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 5221 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 5281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 5341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 5461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 5521 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 5581 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 5641 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 5701 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 5761 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 5821 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 5881 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 5941 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6001 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6061 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6121 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6181 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6241 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6301 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6361 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 6421 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 6481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 6541 gccacctg //