LOCUS V025101 6503 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025101 VERSION V025101 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..2712 /label="OR6Q1(NM_001005186)" /note="OR6Q1(NM_001005186)" /gene="OR6Q1" CDS 2719..2739 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2740..3453 /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 3454..3477 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3496..3519 /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 3591..3646 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4006..4022) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4030..4046 /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(4054..4084) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4099..4120 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4179..4374 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4380..4514 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(4753..5341) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5512..6372) /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(6373..6477) /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 gatgcaacca tataccaaaa actggaccca ggtaactgaa 1801 tttgtcatga tgggctttgc tggcatccat gaagcacacc tcctcttctt catactcttc 1861 ctcaccatgt acctgttcac cttggtggag aatttggcca tcattttagt ggtgggtttg 1921 gaccaccgac tacggagacc catgtatttc ttcctgacac acttgtcctg ccttgaaatc 1981 tggtacactt ctgttacagt gcccaagatg ctggctggtt ttattggggt ggatggtggc 2041 aagaatatct cttatgctga ttgcctatcc cagctcttca tcttcacctt tcttggggca 2101 actgagtgtt tcctactggc tgccatggcc tatgatcgtt atgtggccat ttgtatgcct 2161 ctccactatg gggcttttgt gtcctggggc acctgcatcc gtctggcagc tgcctgttgg 2221 ctggtaggtt tcctcacacc catcttgcca atctacctct tgtctcagct aacattttat 2281 ggcccaaatg tcattgacca tttctcctgt gatgcctcac ccttgctagc cttgtcgtgc 2341 tcagatgtca cttggaagga gactgtggat ttcctggtgt ctctggctgt gctactggcc 2401 tcctctatgg tcattgctgt gtcctatggc aacatcgtct ggacactgct gcacatccgc 2461 tcagctgctg agcgctggaa ggccttctct acctgtgcag ctcacctgac tgtggtgagc 2521 ctcttctatg gcactctttt ctttatgtat gtccagacca aggtgacctc ctccatcaac 2581 ttcaacaagg tggtatctgt cttctactct gttgtcacgc ccatgctcaa tcctctcatc 2641 tacagtctta ggaacaagga agtgaaggga gctctgggtc gagtcttttc tctcaacttt 2701 tggaagggac aggaattcga aaacctgtac ttccaatccg tgagcaaggg cgaggagctg 2761 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 2821 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 2881 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 2941 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 3001 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 3061 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 3121 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 3181 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 3241 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 3301 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 3361 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 3421 gggatcactc tcggcatgga cgagctgtac aagtggagcc acccgcagtt cgagaagggc 3481 ggcggcagca gcgcctggag ccacccgcag ttcgagaagt gaagatcttt ttccctctgc 3541 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa 3601 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata 3661 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata 3721 tgcccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa 3781 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt 3841 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt 3901 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc 3961 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc 4021 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4081 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4141 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc 4201 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 4261 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 4321 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta 4381 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4441 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4501 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 4741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 4801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 4861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 4921 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta 4981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5161 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 5221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 5341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 5401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 5461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 5521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 5581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 5641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 5701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 5761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 5821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 5881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 5941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 6001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 6061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 6241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 6301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 6361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 6421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 6481 atttccccga aaagtgccac ctg //