LOCUS V025066 6468 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025066 VERSION V025066 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 1696..1719 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 1720..1743 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 1744..1815 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" CDS 1816..1836 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 1852..3444 /label="PAF1(NM_019088)" /note="PAF1(NM_019088)" /gene="PAF1" polyA_signal 3555..3610 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(3971..3987) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3995..4011 /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(4019..4049) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4064..4085 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4144..4339 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4345..4479 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(4718..5306) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5477..6337) /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(6338..6442) /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 ctagctggag ccacccgcag ttcgagaagt ggagccaccc gcagttcgag 1741 aaggactaca aggacgatga tgacaaggac tacaaggacg atgatgacaa ggactacaag 1801 gacgatgatg acaaggaaaa cctgtacttc caatccgcga tcgctctcga gatggcgccc 1861 accatccaga cccaggccca gcgggaggat ggccacaggc ccaattccca ccggactctg 1921 cctgagaggt ctggagtggt ctgccgagtc aagtactgca atagcctccc tgatatcccc 1981 ttcgacccca agttcatcac ctaccccttc gaccagaaca ggttcgtcca gtacaaagcc 2041 acttccttgg agaaacagca caaacatgac ctcctgactg agccagacct gggggtcacc 2101 atcgatctca tcaatcctga cacctaccgc atcgacccca atgttcttct agatccagct 2161 gatgagaaac ttttggaaga ggagattcag gcccccacca gctccaagag atcccagcag 2221 cacgcgaagg tggtgccatg gatgcgaaag acagagtaca tctccactga gttcaaccgt 2281 tatggcatct ccaatgagaa gcctgaggtc aagattgggg tttctgtgaa gcagcagttt 2341 accgaggaag aaatatacaa agacagggat agccagatca cagccattga gaagactttt 2401 gaggatgccc agaaatcaat ctcacagcat tacagcaaac cccgagtcac accggtggag 2461 gtcatgcctg tcttcccaga ctttaagatg tggatcaatc catgtgctca ggtgatcttt 2521 gactcagacc cagcccccaa ggacacgagt ggtgcagctg cgttggagat gatgtctcag 2581 gccatgatta ggggcatgat ggatgaggaa gggaaccagt ttgtggccta tttcctgcct 2641 gtagaagaga cgttgaagaa acgaaagcgg gaccaggagg aggagatgga ctatgcacca 2701 gatgatgtgt atgactacaa aattgctcgg gagtacaact ggaacgtgaa gaacaaagct 2761 agcaagggct atgaggaaaa ctacttcttc atcttccgag agggtgacgg ggtttactac 2821 aatgagttgg aaaccagggt ccgccttagt aagcgccggg ccaaggctgg ggttcagtca 2881 ggcaccaacg ccctgcttgt ggtcaaacat cgggacatga atgagaagga actggaagct 2941 caggaggcac ggaaggccca gctagaaaac cacgaaccgg aggaggaaga ggaagaggag 3001 atggagacag aagagaaaga agctgggggc tcagatgagg agcaggagaa gggcagcagc 3061 agtgagaagg agggcagtga agatgagcac tcgggcagcg agagtgaacg ggaggaaggt 3121 gacagggacg aggccagtga caagagtggc agtggtgagg acgagagcag cgaggatgag 3181 gcccgggctg cccgtgacaa agaggagatc tttggcagtg atgctgattc tgaggacgat 3241 gccgactctg atgatgagga cagaggacag gcccaaggtg gcagtgacaa tgattcagac 3301 agcggcagca atgggggtgg ccagcggagc cggagccaca gccgcagcgc cagtcccttc 3361 cccagtggca gcgagcactc ggcccaggag gatggcagtg aagctgcagc ttctgattcc 3421 agtgaagctg atagtgacag tgactgaggt accaccggta cgcgtgaatt cgcggccgca 3481 tctagaagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca 3541 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt 3601 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta 3661 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct 3721 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa 3781 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac 3841 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact 3901 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg 3961 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4021 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4081 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga 4141 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 4201 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 4261 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 4321 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa 4381 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4441 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc 4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac 4921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 6001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 6061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 6121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 6181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 6301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 6361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 6421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg //