LOCUS V020144 9224 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020144 VERSION V020144 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..3904 /label="PLD1(NM_002662)" /note="PLD1(NM_002662)" /gene="PLD1" CDS 3911..3976 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4000..4588 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4654..4702 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4904..5332 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5346..5675 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5742..6536 /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 6712..6833 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6882..6898) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6906..6922 /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(6930..6960) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6975..6996 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7284..7872) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8043..8903) /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(8904..9008) /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 ccatgtcact gaaaaacgag ccacgggtaa atacctctgc 721 actgcagaaa attgctgctg acatgagtaa tatcatagaa aatctggaca cgcgggaact 781 ccactttgag ggagaggagg tagactacga cgtgtctccc agcgatccca agatacaaga 841 agtgtatatc cctttctctg ctatttataa cactcaagga tttaaggagc ctaatataca 901 gacgtatctc tccggctgtc caataaaagc acaagttctg gaagtggaac gcttcacatc 961 tacaacaagg gtaccaagta ttaatcttta cactattgaa ttaacacatg gggaatttaa 1021 atggcaagtt aagaggaaat tcaagcattt tcaagaattt cacagagagc tgctcaagta 1081 caaagccttt atccgcatcc ccattcccac tagaagacac acgtttagga ggcaaaacgt 1141 cagagaggag cctcgagaga tgcccagttt gccccgttca tctgaaaaca tgataagaga 1201 agaacaattc cttggtagaa gaaaacaact ggaagattac ttgacaaaga tactaaaaat 1261 gcccatgtat agaaactatc atgccacaac agagtttctt gatataagcc agctgtcttt 1321 catccatgat ttgggaccaa agggcataga aggtatgata atgaaaagat ctggaggaca 1381 cagaatacca ggcttgaatt gctgtggtca gggaagagcc tgctacagat ggtcaaaaag 1441 atggttaata gtgaaagatt cctttttatt gtatatgaaa ccagacagcg gtgccattgc 1501 cttcgtcctg ctggtagaca aagaattcaa aattaaggtg gggaagaagg agacagaaac 1561 gaaatatgga atccgaattg ataatctttc aaggacactt attttaaaat gcaacagcta 1621 tagacatgct cggtggtggg gaggggctat agaagaattc atccagaaac atggcaccaa 1681 ctttctcaaa gatcatcgat ttgggtcata tgctgctatc caagagaatg ctttagctaa 1741 atggtatgtt aatgccaaag gatattttga agatgtggca aatgcaatgg aagaggcaaa 1801 tgaagagatt tttatcacag actggtggct gagtccagaa atcttcctga aacgcccagt 1861 ggttgaggga aatcgttgga ggttggactg cattcttaaa cgaaaagcac aacaaggagt 1921 gaggatcttc ataatgctct acaaagaggt ggaactcgct cttggcatca atagtgaata 1981 caccaagagg actttgatgc gtctacatcc caacataaag gtgatgagac acccggatca 2041 tgtgtcatcc accgtctatt tgtgggctca ccatgagaag cttgtcatca ttgaccaatc 2101 ggtggccttt gtgggaggga ttgacctggc ctatggaagg tgggacgaca atgagcacag 2161 actcacagac gtgggcagtg tgaagcgggt cacttcagga ccgtctctgg gttccctccc 2221 acctgccgca atggagtcta tggaatcctt aagactcaaa gataaaaatg agcctgttca 2281 aaacctaccc atccagaaga gtattgatga tgtggattca aaactgaaag gaataggaaa 2341 gccaagaaag ttctccaaat ttagtctcta caagcagctc cacaggcacc acctgcacga 2401 cgcagatagc atcagcagca ttgacagcac ctccagttat tttaatcact atagaagtca 2461 tcacaattta atccatggtt taaaacccca cttcaaactc tttcacccgt ccagtgagtc 2521 tgagcaagga ctcactagac ctcatgctga taccgggtcc atccgtagtt tacagacagg 2581 tgtgggagag ctgcatgggg aaaccagatt ctggcatgga aaggactact gcaatttcgt 2641 cttcaaagac tgggttcaac ttgataaacc ttttgctgat ttcattgaca ggtactccac 2701 gccccggatg ccctggcatg acattgcctc tgcagtccac gggaaggcgg ctcgtgatgt 2761 ggcacgtcac ttcatccagc gctggaactt cacaaaaatt atgaaatcaa aatatcggtc 2821 cctttcttat ccttttctgc ttccaaagtc tcaaacaaca gcccatgagt tgagatatca 2881 agtgcctggg tctgtccatg ctaacgtaca gttgctccgc tctgctgctg attggtctgc 2941 tggtataaag taccatgaag agtccatcca cgccgcttac gtccatgtga tagagaacag 3001 caggcactat atctatatcg aaaaccagtt tttcataagc tgtgctgatg acaaagttgt 3061 gttcaacaag ataggcgatg ccattgccca gaggatcctg aaagctcaca gggaaaacca 3121 gaaataccgg gtatatgtcg tgataccact tctgccaggg ttcgaaggag acatttcaac 3181 cggcggagga aatgctctac aggcaatcat gcacttcaac tacagaacca tgtgcagagg 3241 agaaaattcc atccttggac agttaaaagc agagcttggt aatcagtgga taaattacat 3301 atcattctgt ggtcttagaa cacatgcaga gctcgaagga aacctagtaa ctgagcttat 3361 ctatgtccac agcaagttgt taattgctga tgataacact gttattattg gctctgccaa 3421 cataaatgac cgcagcatgc tgggaaagcg tgacagtgaa atggctgtca ttgtgcaaga 3481 tacagagact gttccttcag taatggatgg aaaagagtac caagctggcc ggtttgcccg 3541 aggacttcgg ctacagtgct ttagggttgt ccttggctat cttgatgacc caagtgagga 3601 cattcaggat ccagtgagtg acaaattctt caaggaggtg tgggtttcaa cagcagctcg 3661 aaatgctaca atttatgaca aggttttccg gtgccttccc aatgatgaag tacacaattt 3721 aattcagctg agagacttta taaacaagcc cgtattagct aaggaagatc ccattcgagc 3781 tgaggaggaa ctgaagaaga tccgtggatt tttggtgcaa ttcccctttt atttcttgtc 3841 tgaagaaagc ctactgcctt ctgttgggac caaagaggcc atagtgccca tggaggtttg 3901 gactctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 3961 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 4021 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4081 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4141 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 4201 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 4261 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 4321 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 4381 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 4441 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 4501 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 4561 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 4621 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 4681 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 4741 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4801 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 4861 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 4921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 4981 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 5041 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 5101 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 5161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 5221 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 5281 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 5341 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 5401 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 5461 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 5521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 5581 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 5641 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 5701 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 5761 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 5821 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 5881 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 5941 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 6001 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 6061 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 6121 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 6181 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 6241 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 6301 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 6361 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 6421 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 6481 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 6541 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 6601 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 6661 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 6721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 6781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 6841 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 6901 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6961 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 7021 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 7081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 7321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 7381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 7441 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 7501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 7561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7681 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 7741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7921 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7981 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 8041 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 8101 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 8161 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 8221 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 8281 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 8341 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 8401 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 8461 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 8521 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 8581 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8641 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8701 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8761 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8821 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8881 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8941 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 9001 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 9061 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 9121 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 9181 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //