LOCUS V019523 8186 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019523 VERSION V019523 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..2866 /label="PCCA(NM_000282)" /note="PCCA(NM_000282)" /gene="PCCA" CDS 2873..2938 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2962..3550 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3616..3664 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3866..4294 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4308..4637 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4704..5498 /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 5674..5795 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5844..5860) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5868..5884 /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(5892..5922) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5937..5958 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6246..6834) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7005..7865) /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(7866..7970) /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 ccatggcggg gttctgggtc gggacagcac cgctggtcgc 721 tgccggacgg cgtgggcggt ggccgccgca gcagctgatg ctgagcgcgg cgctgcggac 781 cctgaagcat gttctgtact attcaagaca gtgcttaatg gtgtcccgta atcttggttc 841 agtgggatat gatcctaatg aaaaaacttt tgataaaatt cttgttgcta atagaggaga 901 aattgcatgt cgggttatta gaacttgcaa gaagatgggc attaagacag ttgccatcca 961 cagtgatgtt gatgctagtt ctgttcatgt gaaaatggcg gatgaggctg tctgtgttgg 1021 cccagctccc accagtaaaa gctacctcaa catggatgcc atcatggaag ccattaagaa 1081 aaccagggcc caagctgtac atccaggtta tggattcctt tcagaaaaca aagaatttgc 1141 cagatgtttg gcagcagaag atgtcgtttt cattggacct gacacacatg ctattcaagc 1201 catgggcgac aagattgaaa gcaaattatt agctaagaaa gcagaggtta atacaatccc 1261 tggctttgat ggagtagtca aggatgcaga agaagctgtc agaattgcaa gggaaattgg 1321 ctaccctgtc atgatcaagg cctcagcagg tggtggtggg aaaggcatgc gcattgcttg 1381 ggatgatgaa gagaccaggg atggttttag attgtcatct caagaagctg cttctagttt 1441 tggcgatgat agactactaa tagaaaaatt tattgataat cctcgtcata tagaaatcca 1501 ggttctaggt gataaacatg ggaatgcttt atggcttaat gaaagagagt gctcaattca 1561 gagaagaaat cagaaggtgg tggaggaagc accaagcatt tttttggatg cggagactcg 1621 aagagcgatg ggagaacaag ctgtagctct tgccagagca gtaaaatatt cctctgctgg 1681 gaccgtggag ttccttgtgg actctaagaa gaatttttat ttcttggaaa tgaatacaag 1741 actccaggtt gagcatcctg tcacagaatg cattactggc ctggacctag tccaggaaat 1801 gatccgtgtt gctaagggct accctctcag gcacaaacaa gctgatattc gcatcaacgg 1861 ctgggcagtt gaatgtcggg tttatgctga ggacccctac aagtcttttg gtttaccatc 1921 tattgggaga ttgtctcagt accaagaacc gttacatcta cctggtgtcc gagtggacag 1981 tggcatccaa ccaggaagtg atattagcat ttattatgat cctatgattt caaaactaat 2041 cacatatggc tctgatagaa ctgaggcact gaagagaatg gcagatgcac tggataacta 2101 tgttattcga ggtgttacac ataatattgc attacttcga gaggtgataa tcaactcacg 2161 ctttgtaaaa ggagacatca gcactaaatt tctctccgat gtgtatcctg atggcttcaa 2221 aggacacatg ctaaccaaga gtgagaagaa ccagttattg gcaatagcat catcattgtt 2281 tgtggcattc cagttaagag cacaacattt tcaagaaaat tcaagaatgc ctgttattaa 2341 accagacata gccaactggg agctctcagt aaaattgcat gataaagttc ataccgtagt 2401 agcatcaaac aatgggtcag tgttctcggt ggaagttgat gggtcgaaac taaatgtgac 2461 cagcacgtgg aacctggctt cgcccttatt gtctgtcagc gttgatggca ctcagaggac 2521 tgtccagtgt ctttctcgag aagcaggtgg aaacatgagc attcagtttc ttggtacagt 2581 gtacaaggtg aatatcttaa ccagacttgc cgcagaattg aacaaattta tgctggaaaa 2641 agtgactgag gacacaagca gtgttctgcg ttccccgatg cccggagtgg tggtggccgt 2701 ctctgtcaag cctggagacg cggtagcaga aggtcaagaa atttgtgtga ttgaagccat 2761 gaaaatgcag aatagtatga cagctgggaa aactggcacg gtgaaatctg tgcactgtca 2821 agctggagac acagttggag aaggggatct gctcgtggag ctggaactcg aggactacaa 2881 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2941 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact 3001 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 3061 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 3121 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 3181 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 3241 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 3301 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 3361 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 3421 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 3481 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 3541 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 3601 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt 3661 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 3721 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 3781 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc 3841 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3901 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3961 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 4021 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 4081 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4141 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4201 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa 4261 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4381 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4441 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 4501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4561 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4621 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 4681 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 4741 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 4801 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 4861 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 4921 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4981 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 5041 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 5101 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5161 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 5221 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 5281 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 5341 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 5401 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 5461 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa 5521 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 5581 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 5641 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 5701 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 5761 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5821 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5881 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5941 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 6001 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 6061 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 6121 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 6181 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 6241 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6301 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6361 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6421 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6481 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6541 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6601 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 6661 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 6721 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6781 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6841 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6901 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6961 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 7021 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 7081 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 7141 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 7201 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 7261 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 7321 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 7381 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 7441 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 7501 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 7561 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 7621 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 7681 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 7741 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 7801 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7861 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7921 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7981 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc 8041 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 8101 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 8161 acaattgcat gaagaatctg cttagg //