LOCUS V024994 6650 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024994 VERSION V024994 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..2859 /label="HTR1F(NM_000866)" /note="HTR1F(NM_000866)" /gene="HTR1F" CDS 2866..2886 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2887..3600 /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 3601..3624 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3643..3666 /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 3738..3793 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4153..4169) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4177..4193 /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(4201..4231) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4246..4267 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4326..4521 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4527..4661 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(4900..5488) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5659..6519) /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(6520..6624) /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 gatggatttc ttaaattcat ctgatcaaaa cttgacctca 1801 gaggaactgt taaacagaat gccatccaaa attctggtgt ccctcactct gtctgggctg 1861 gcactgatga caacaactat caactccctt gtgatcgctg caattattgt gacccggaag 1921 ctgcaccatc cagccaatta tttaatttgt tcccttgcag tcacagattt tcttgtggct 1981 gtcctggtga tgcccttcag cattgtgtat attgtgagag agagctggat tatggggcaa 2041 gtggtctgtg acatttggct gagtgttgac attacctgct gcacgtgctc catcttgcat 2101 ctctcagcta tagctttgga tcggtatcga gcaatcacag atgctgttga gtatgccagg 2161 aaaaggactc caaagcatgc tggcattatg attacaatag tttggattat atctgttttt 2221 atctctatgc ctcctctatt ctggaggcac caaggaacta gcagagatga tgaatgcatc 2281 atcaagcacg accacattgt ttccaccatt tactcaacat ttggagcttt ctacatccca 2341 ctggcattga ttttgatcct ttactacaaa atatatagag cagcaaagac attataccac 2401 aagagacaag caagtaggat tgcaaaggag gaggtgaatg gccaagtcct tttggagagt 2461 ggtgagaaaa gcactaaatc agtttccaca tcctatgtac tagaaaagtc tttatctgac 2521 ccatcaacag actttgataa aattcatagc acagtgagaa gtctcaggtc tgaattcaag 2581 catgagaaat cttggagaag gcaaaagatc tcaggtacaa gagaacggaa agcagccact 2641 accctgggat taatcttggg tgcatttgta atatgttggc ttcctttttt tgtaaaagaa 2701 ttagttgtta atgtctgtga caaatgtaaa atttctgaag aaatgtccaa ttttttggca 2761 tggcttgggt atctcaattc ccttataaat ccactgattt acacaatctt taatgaagac 2821 ttcaagaaag cattccaaaa gcttgtgcga tgtcgatgtg aattcgaaaa cctgtacttc 2881 caatccgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 2941 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 3001 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 3061 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 3121 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 3181 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 3241 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 3301 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 3361 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 3421 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 3481 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 3541 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 3601 tggagccacc cgcagttcga gaagggcggc ggcagcagcg cctggagcca cccgcagttc 3661 gagaagtgaa gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga 3721 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt 3781 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga 3841 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggtgg 3901 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga 3961 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta 4021 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta 4081 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt 4141 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4201 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4261 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg 4321 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 4381 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 4441 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 4501 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat 4561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4621 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg 4681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 4741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 5101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5341 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg //