LOCUS V025076 6890 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025076 VERSION V025076 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..3099 /label="DRD1(NM_000794)" /note="DRD1(NM_000794)" /gene="DRD1" CDS 3106..3126 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 3127..3840 /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 3841..3864 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3883..3906 /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 3978..4033 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4393..4409) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4417..4433 /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(4441..4471) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4486..4507 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4566..4761 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4767..4901 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(5140..5728) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5899..6759) /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(6760..6864) /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 gatgaggact ctgaacacct ctgccatgga cgggactggg 1801 ctggtggtgg agagggactt ctctgttcgt atcctcactg cctgtttcct gtcgctgctc 1861 atcctgtcca cgctcctggg gaacacgctg gtctgtgctg ccgttatcag gttccgacac 1921 ctgcggtcca aggtgaccaa cttctttgtc atctccttgg ctgtgtcaga tctcttggtg 1981 gccgtcctgg tcatgccctg gaaggcagtg gctgagattg ctggcttctg gccctttggg 2041 tccttctgta acatctgggt ggcctttgac atcatgtgct ccactgcatc catcctcaac 2101 ctctgtgtga tcagcgtgga caggtattgg gctatctcca gccctttccg gtatgagaga 2161 aagatgaccc ccaaggcagc cttcatcctg atcagtgtgg catggacctt gtctgtactc 2221 atctccttca tcccagtgca gctcagctgg cacaaggcaa aacccacaag cccctctgat 2281 ggaaatgcca cttccctggc tgagaccata gacaactgtg actccagcct cagcaggaca 2341 tatgccatct catcctctgt aataagcttt tacatccctg tggccatcat gattgtcacc 2401 tacaccagga tctacaggat tgctcagaaa caaatacggc gcattgcggc cttggagagg 2461 gcagcagtcc acgccaagaa ttgccagacc accacaggta atggaaagcc tgtcgaatgt 2521 tctcaaccgg aaagttcttt taagatgtcc ttcaaaagag aaactaaagt cctgaagact 2581 ctgtcggtga tcatgggtgt gtttgtgtgc tgttggctac ctttcttcat cttgaactgc 2641 attttgccct tctgtgggtc tggggagacg cagcccttct gcattgattc caacaccttt 2701 gacgtgtttg tgtggtttgg gtgggctaat tcatccttga accccatcat ttatgccttt 2761 aatgctgatt ttcggaaggc attttcaacc ctcttaggat gctacagact ttgccctgcg 2821 acgaataatg ccatagagac ggtgagtatc aataacaatg gggccgcgat gttttccagc 2881 catcatgagc cacgaggctc catctccaag gagtgcaatc tggtttacct gatcccacat 2941 gctgtgggct cctctgagga cctgaaaaag gaggaggcag ctggcatcgc cagacccttg 3001 gagaagctgt ccccagccct atcagtcata ttggactatg acactgacgt ctctctggag 3061 aagatccaac ccatcacaca aaacggtcag cacccaaccg aattcgaaaa cctgtacttc 3121 caatccgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 3181 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 3241 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 3301 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 3361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 3421 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 3481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 3541 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 3601 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 3661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 3721 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 3781 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 3841 tggagccacc cgcagttcga gaagggcggc ggcagcagcg cctggagcca cccgcagttc 3901 gagaagtgaa gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga 3961 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt 4021 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga 4081 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggtgg 4141 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga 4201 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta 4261 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta 4321 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt 4381 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4441 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4501 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg 4561 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 4621 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 4681 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 4741 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat 4801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4861 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg 4921 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 4981 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 5341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5581 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5821 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5881 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5941 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 6001 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 6061 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 6121 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 6181 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6241 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6301 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6361 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6421 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6481 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6541 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6601 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6661 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6721 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6781 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6841 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg //