LOCUS V019234 7394 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019234 VERSION V019234 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..2074 /label="SCG3(NM_013243)" /note="SCG3(NM_013243)" /gene="SCG3" CDS 2081..2146 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2170..2758 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2824..2872 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3074..3502 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3516..3845 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3912..4706 /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 4882..5003 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5052..5068) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5076..5092 /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(5100..5130) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5145..5166 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5454..6042) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6213..7073) /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(7074..7178) /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 atggggttcc tcgggaccgg cacttggatt ctggtgttag tgctcccgat 721 tcaagctttc cccaaacctg gaggaagcca agacaaatct ctacataata gagaattaag 781 tgcagaaaga cctttgaatg aacagattgc tgaagcagaa gaagacaaga ttaaaaaaac 841 atatcctcca gaaaacaagc caggtcagag caactattct tttgttgata acttgaacct 901 gctaaaggca ataacagaaa aggaaaaaat tgagaaagaa agacaatcta taagaagctc 961 cccacttgat aataagttga atgtggaaga tgttgattca accaagaatc gaaaactgat 1021 cgatgattat gactctacta agagtggatt ggatcataaa tttcaagatg atccagatgg 1081 tcttcatcaa ctagacggga ctcctttaac cgctgaagac attgtccata aaatcgctgc 1141 caggatttat gaagaaaatg acagagccgt gtttgacaag attgtttcta aactacttaa 1201 tctcggcctt atcacagaaa gccaagcaca tacactggaa gatgaagtag cagaggtttt 1261 acaaaaatta atctcaaagg aagccaacaa ttatgaggag gatcccaata agcccacaag 1321 ctggactgag aatcaggctg gaaaaatacc agagaaagtg actccaatgg cagcaattca 1381 agatggtctt gctaagggag aaaacgatga aacagtatct aacacattaa ccttgacaaa 1441 tggcttggaa aggagaacta aaacctacag tgaagacaac tttgaggaac tccaatattt 1501 cccaaatttc tatgcgctac tgaaaagtat tgattcagaa aaagaagcaa aagagaaaga 1561 aacactgatt actatcatga aaacactgat tgactttgtg aagatgatgg tgaaatatgg 1621 aacaatatct ccagaagaag gtgtttccta ccttgaaaac ttggatgaaa tgattgctct 1681 tcagaccaaa aacaagctag aaaaaaatgc tactgacaat ataagcaagc ttttcccagc 1741 accatcagag aagagtcatg aagaaacaga cagtaccaag gaagaagcag ctaagatgga 1801 aaaggaatat ggaagcttga aggattccac aaaagatgat aactccaacc caggaggaaa 1861 gacagatgaa cccaaaggaa aaacagaagc ctatttggaa gccatcagaa aaaatattga 1921 atggttgaag aaacatgaca aaaagggaaa taaagaagat tatgaccttt caaagatgag 1981 agacttcatc aataaacaag ctgatgctta tgtggagaaa ggcatccttg acaaggaaga 2041 agccgaggcc atcaagcgca tttatagcag cctgctcgag gactacaaag accatgacgg 2101 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 2161 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 2221 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 2281 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 2341 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 2401 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 2461 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 2521 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 2581 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 2641 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 2701 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 2761 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 2821 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 2881 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 2941 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3001 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 3061 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 3241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3421 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 3481 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3781 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3841 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3901 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3961 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4021 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4081 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4141 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4201 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4261 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4321 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4381 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4441 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4501 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4561 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4621 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4681 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 4741 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 4801 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4861 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4981 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 5041 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 5221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 5281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 5341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 5401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 5521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 5581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5641 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 5821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5881 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7201 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 7261 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 7321 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 7381 agaatctgct tagg //