LOCUS V022267 7289 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022267 VERSION V022267 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..1969 /label="ACTL6A(NM_004301)" /note="ACTL6A(NM_004301)" /gene="ACTL6A" CDS 1976..2041 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2065..2653 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2719..2767 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2969..3397 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3411..3740 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3807..4601 /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 4777..4898 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4947..4963) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4971..4987 /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(4995..5025) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5040..5061 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5349..5937) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6108..6968) /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(6969..7073) /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 ccatgagcgg cggcgtgtac gggggagatg aagttggagc 721 ccttgttttt gacattggat cctatactgt gagagctggt tatgctggtg aggactgccc 781 caaggtggat tttcctacag ctattggtat ggtggtagaa agagatgacg gaagcacatt 841 aatggaaata gatggcgata aaggcaaaca aggcggtccc acctactaca tagatactaa 901 tgctctgcgt gttccgaggg agaatatgga ggccatttca cctctaaaaa atgggatggt 961 tgaagactgg gatagtttcc aagctatttt ggatcatacc tacaaaatgc atgtcaaatc 1021 agaagccagt ctccatcctg ttctcatgtc agaggcaccg tggaatacta gagcaaagag 1081 agagaaactg acagagttaa tgtttgaaca ctacaacatc cctgccttct tcctttgcaa 1141 aactgcagtt ttgacagcat ttgctaatgg tcgttctact gggctgattt tggacagtgg 1201 agccactcat accactgcaa ttccagtcca cgatggctat gtccttcaac aaggcattgt 1261 gaaatcccct cttgctggag actttattac tatgcagtgc agagaactct tccaagaaat 1321 gaatattgaa ttggttcctc catatatgat tgcatcaaaa gaagctgttc gtgaaggatc 1381 tccagcaaac tggaaaagaa aagagaagtt gcctcaggtt acgaggtctt ggcacaatta 1441 tatgtgtaat tgtgttatcc aggattttca agcttcggta cttcaagtgt cagattcaac 1501 ttatgatgaa caagtggctg cacagatgcc aactgttcat tatgaattcc ccaatggcta 1561 caattgtgat tttggtgcag agcggctaaa gattccagaa ggattatttg acccttccaa 1621 tgtaaagggg ttatcaggaa acacaatgtt aggagtcagt catgttgtca ccacaagtgt 1681 tgggatgtgt gatattgaca tcagaccagg tctctatggc agtgtaatag tggcaggagg 1741 aaacacacta atacagagtt ttactgacag gttgaataga gagctgtctc agaaaactcc 1801 tccaagtatg cggttgaaat tgattgcaaa taatacaaca gtggaacgga ggtttagctc 1861 atggattggc ggctccattc tagcctcttt gggtaccttt caacagatgt ggatttccaa 1921 gcaagaatat gaagaaggag ggaagcagtg tgtagaaaga aaatgccctc tcgaggacta 1981 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa 2041 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg 2101 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 2161 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 2221 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 2281 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 2341 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 2401 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 2461 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 2521 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 2581 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 2641 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg 2701 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg 2761 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 2821 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 2881 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg 2941 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 3001 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 3061 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 3121 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3181 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3241 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3301 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta 3361 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3421 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3481 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3541 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3661 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 3721 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 3781 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3961 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 4321 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc 4621 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4681 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4741 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 4801 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 4861 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 4921 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 4981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 5041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 5101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 5161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 5221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 5281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 5341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 5401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 5461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 5521 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 5581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 5641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 5701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 5761 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 5821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 5881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 5941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 6001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 6721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 6781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 6841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 6901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 6961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 7021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 7081 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga 7141 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 7201 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 7261 ccgacaattg catgaagaat ctgcttagg //