LOCUS V019490 7100 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019490 VERSION V019490 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..1780 /label="Zdhhc2(NM_178395)" /note="Zdhhc2(NM_178395)" /gene="Zdhhc2" CDS 1787..1852 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1876..2464 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2530..2578 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2780..3208 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3222..3551 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3618..4412 /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 4588..4709 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4758..4774) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4782..4798 /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(4806..4836) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4851..4872 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5160..5748) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5919..6779) /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(6780..6884) /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 ccatggcgcc ctcgggctcg ggcggcgtga ggcggcggtg 721 ccggcgggtg ctctactgga tcccggtggt gttcatcagc ttgctgctgg gctggtccta 781 ctacgcctac gccatccagc tgtgcatagt gtccatggaa aacattggtg aacaagttgt 841 gtgcctcatg gcttatcatc tactttttgc aatgtttgtc tggtcatact ggaaaaccat 901 ttttacattg cccatgaatc cttcaaaaga attccatctc tcttatgcag agaaagaatt 961 gctggagaga gagccaagag gagaagccca tcaggaagtt ctgaggcgag cagccaaaga 1021 ccttcccatc tacaccagga ccatgtccgg cgcaatccga tattgtgaca gatgccaact 1081 tataaaacca gaccgctgtc atcattgttc cgtctgtgat aaatgtattt tgaagatgga 1141 tcatcattgc ccatgggtga acaattgtgt tggattttca aactacaaat tcttccttct 1201 tttcttggct tactctctgc tgtactgcct tttcattgct gctaccgatt tacagtattt 1261 tatcagattt tggacaaatg gtctgcctga tactcaagcc aagttccata ttatgttttt 1321 attctttgct gcagctatgt tttctgtcag cttgtcctct ctgtttggtt atcattgctg 1381 gctagtcagc aaaaataaat ctactttaga ggcattcaga aatccagtat ttagacacgg 1441 aacagataag aacggattca gcttgggttt cagtaaaaac atgagacaag tgtttggtga 1501 tgagaagaaa tactggctgt taccagtatt ttcaagtcaa ggtgatggct gttcctttcc 1561 aacttgcctt gttaaccagg atcctgaaca accttctact cctgcgggac taaattcaac 1621 agtgaaaaat cctgaaaacc accagtttcc tgcaaagcct ctgagagagt cccagagcca 1681 tctccttaag gattctcaga cctggacaga gagcagcgca aaccctggga agggcaaagc 1741 cggtatgagc aaccctgcat taactatgga gaacgagact ctcgaggact acaaagacca 1801 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg 1861 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 1921 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 1981 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 2041 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 2101 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 2161 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 2221 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 2281 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 2341 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 2401 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 2461 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 2521 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat 2581 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 2641 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 2701 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg 2761 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 2821 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 2881 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc 2941 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 3001 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 3061 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 3121 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag 3181 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 3421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 3481 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 3541 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 3601 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 3661 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 3721 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 3781 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 3841 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 3901 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 3961 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 4021 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 4081 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 4141 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 4201 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 4261 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 4321 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 4381 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc 4441 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 4501 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 4561 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 4621 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 4681 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 4741 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 4801 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 4861 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 4921 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 4981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 5041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 5101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 5161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 5221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 5281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 5341 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 5401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 5461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 5521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 5581 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 5641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 5701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 5761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 5821 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 5881 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 5941 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 6001 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 6061 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 6121 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 6181 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 6241 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 6301 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 6361 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 6421 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 6481 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 6541 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 6601 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 6661 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 6721 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 6781 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 6841 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 6901 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta 6961 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg 7021 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 7081 gcatgaagaa tctgcttagg //