LOCUS V019142 7088 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019142 VERSION V019142 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..1735 /label="PRKACB(NM_002731)" /note="PRKACB(NM_002731)" /gene="PRKACB" CDS 1775..1840 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1864..2452 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2518..2566 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2768..3196 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3210..3539 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3606..4400 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4576..4697 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4746..4762) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4770..4786 /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(4794..4824) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4839..4860 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5148..5736) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5907..6767) /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(6768..6872) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatggggaa cgcggcgacc gccaagaaag gcagcgaggt 721 ggagagcgtg aaagagtttc tagccaaagc caaagaagac tttttgaaaa aatgggagaa 781 tccaactcag aataatgccg gacttgaaga ttttgaaagg aaaaaaaccc ttggaacagg 841 ttcatttgga agagtcatgt tggtaaaaca caaagccact gaacagtatt atgccatgaa 901 gatcttagat aagcagaagg ttgttaaact gaagcaaata gagcatactt tgaatgagaa 961 aagaatatta caggcagtga attttccttt ccttgttcga ctggagtatg cttttaagga 1021 taattctaat ttatacatgg ttatggaata tgtccctggg ggtgaaatgt tttcacatct 1081 aagaagaatt ggaaggttca gtgagcccca tgcacggttc tatgcagctc agatagtgct 1141 aacattcgag tacctccatt cactagacct catctacaga gatctaaaac ctgaaaatct 1201 cttaattgac catcaaggct atatccaggt cacagacttt gggtttgcca aaagagttaa 1261 aggcagaact tggacattat gtggaactcc agagtatttg gctccagaaa taattctcag 1321 caagggctac aataaggcag tggattggtg ggcattagga gtgctaatct atgaaatggc 1381 agctggctat cccccattct ttgcagacca accaattcag atttatgaaa agattgtttc 1441 tggaaaggtc cgattcccat cccacttcag ttcagatctc aaggaccttc tacggaacct 1501 gctgcaggtg gatttgacca agagatttgg aaatctaaag aatggtgtca gtgatataaa 1561 aactcacaag tggtttgcca cgacagattg gattgctatt taccagagga aggttgaagc 1621 tccattcata ccaaagttta gaggctctgg agataccagc aactttgatg actatgaaga 1681 agaagatatc cgtgtctcta taacagaaaa atgtgcaaaa gaatttggtg aatttgaatt 1741 ctgcagatat ccagcacagt ggcggccgct cgaggactac aaagaccatg acggtgatta 1801 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 1861 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 1921 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 1981 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 2041 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 2101 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 2161 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 2221 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 2281 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 2341 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 2401 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 2461 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 2521 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 2581 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2641 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2701 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 2761 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2821 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2881 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 2941 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 3001 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 3061 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 3121 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 3181 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3541 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3601 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3661 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3721 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3781 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3841 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3901 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3961 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 4021 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 4081 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 4141 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 4201 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4261 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4321 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 4381 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 4441 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4501 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4561 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4621 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4681 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4741 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4801 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4861 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4921 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4981 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 5041 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5101 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5161 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5221 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5281 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5341 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5401 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5461 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5521 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5581 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5641 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5701 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5761 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5821 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5881 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5941 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 6001 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 6061 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 6121 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 6181 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6241 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6301 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6361 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6421 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6481 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6541 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6601 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6661 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6721 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6781 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6841 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6901 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 6961 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 7021 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 7081 tgcttagg //