LOCUS V019319 7244 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019319 VERSION V019319 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 671..1924 /label="AVPR1A(NM_000706)" /note="AVPR1A(NM_000706)" /gene="AVPR1A" CDS 1931..1996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2020..2608 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2674..2722 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2924..3352 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3366..3695 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3762..4556 /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 4732..4853 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4902..4918) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4926..4942 /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(4950..4980) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4995..5016 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5304..5892) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6063..6923) /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(6924..7028) /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 taccgccacc atgcgtctct ccgccggtcc cgacgcgggg ccctcgggca actccagccc 721 atggtggcct ctggccaccg gcgctggcaa cacaagccgg gaggccgaag ccctcgggga 781 gggcaacggc ccaccgaggg acgtgcgcaa cgaggagctg gccaaactgg agatcgccgt 841 gctggcggtg actttcgcgg tggccgtgct gggcaacagc agcgtactgc tggctctgca 901 ccggacgccg cgcaagacgt cccgcatgca cctcttcatc cgacacctca gcctggccga 961 cctggccgtg gcattcttcc aggtgctgcc gcaaatgtgc tgggacatca cctaccgctt 1021 ccgcggcccc gactggctgt gccgcgtggt gaagcacctg caggtgttcg gcatgtttgc 1081 gtcggcctac atgctggtag tcatgacagc cgaccgctac atcgcggtgt gccacccgct 1141 caagactctg caacagcccg cgcgccgctc gcgcctcatg atcgcggccg cctgggtgct 1201 gagcttcgtg ctgagcacgc cgcagtactt cgtcttctcc atgatcgagg tgaacaatgt 1261 caccaaggcc cgcgactgct gggccacctt catccagccc tggggttctc gtgcctacgt 1321 gacctggatg acgggcggca tctttgtggc gcccgtggtc atcttgggta cctgctacgg 1381 cttcatctgc tacaacatct ggtgcaacgt ccgcgggaag acggcgtcgc gccagagcaa 1441 gggtgcagag caagcgggtg tggccttcca aaaggggttc ctgctcgcac cctgtgtcag 1501 cagcgtgaag tccatttccc gggccaagat ccgcacggtg aagatgactt ttgtgatcgt 1561 gacggcttac atcgtctgct gggcgccttt cttcatcatc cagatgtggt ctgtctggga 1621 tcccatgtcc gtctggaccg aatcggaaaa ccctaccatc accatcactg cattactggg 1681 ttccttgaat agctgctgta atccctggat atacatgttt tttagtggcc atctccttca 1741 agactgtgtt caaagcttcc catgctgcca aaacatgaag gaaaaattca acaaagaaga 1801 tactgacagt atgagcagaa gacagacttt ttattctaac aatcgaagcc caacaaacag 1861 tacgggtatg tggaaggact cgcctaaatc ttccaagtcc atcaaattca ttcctgtttc 1921 aactctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 1981 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 2041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 2101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 2161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 2221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 2281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 2341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 2401 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 2461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 2521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 2581 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 2641 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 2701 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 2761 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 2821 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 2881 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3061 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3361 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3601 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3661 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3721 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4561 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 4621 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 4681 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 4741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4861 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 4921 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 4981 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5461 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 5521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5701 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 5761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7021 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 7081 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 7141 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 7201 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //