LOCUS V046263 7206 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046263 VERSION V046263 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..1886 /label="Serpinb9g(NM_011455)" /note="Serpinb9g(NM_011455)" /gene="Serpinb9g" CDS 1893..1958 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1982..2570 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2636..2684 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2886..3314 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3328..3657 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3724..4518 /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 4694..4815 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4864..4880) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4888..4904 /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(4912..4942) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4957..4978 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5266..5854) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6025..6885) /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(6886..6990) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga tactctatct caagccaatg 781 gcacctttgc catccatctt ttgaaggtgc tctgtcaaga caacccttca aaaaatgtgt 841 gttattctcc tatgagcatc tcctctgctc tagccatggt tctcttgggt gctaagggag 901 acactgcagt ccagatatgt caggcacttc atttaaaccc agatgaagat gtccatcagg 961 gcttccagtt gcttctccat aacctgaaca agcaaaacaa ccaaaagtac tgccttacta 1021 tggccaacag gctctttgta gaaaatactt gtgaattact tccaacattt aaggagtcct 1081 gtcttaaatt ctaccactct gagatggagc agctctcttt tgccgaagct gccgaggagt 1141 ccaggcaaca cataaacatg tgggtctcca aacagactaa tggtaaaatt ccagacttat 1201 tgtcaaaaga ctcagttaat tcacagacca ggctgattct cgccaatgcc ttatatttcc 1261 atggaacatg gtgtaaacgt tttgaaaaaa ataggaccaa ggaaatgccc tttaagataa 1321 acaagaagga aacaagacca gtgcagatga tgtggcggga agacacatta ttccatgcct 1381 atgtgaagga aatccaggca caagtgctag tgatgcccta tgagggcata gatttgaatt 1441 tcgtggtcct actcccagat gagggtgtgg acatcagcaa ggtggaaaac aatctcactt 1501 ttgagaagtt aacagcctgg accaaaccag agtttatgaa tcgcactgag tttcatgttt 1561 atcttccaaa atttcaacta caagaggatt atgatatgaa ttccctgtta cagcatttgg 1621 gaattttgaa tgtcttcgat ggaagcaagg ctgacttatc aggaatgtct actaaggaaa 1681 acctgtgtct gtccgagttt gctcacaagt gtgtagtaga ggtcaatgaa gaaggaacag 1741 aggctgctgc agcctctgct gtaaaattta tatttttatg ttcaggtcct gatcctgaaa 1801 ctttctgtgc tgaccacccc ttccttttct tcatcatgca cagaacaact aacagcatcc 1861 tgttctgtgg cagattctca tctccaggat ccgactacaa agaccatgac ggtgattata 1921 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga 1981 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2041 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2101 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2161 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2221 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 2281 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 2341 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 2401 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 2461 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 2521 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg 2581 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 2641 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg 2701 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 2761 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 2821 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta 2881 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2941 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3001 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 3061 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3121 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3181 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3241 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3301 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3361 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3481 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3541 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3601 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3661 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3721 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3781 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3841 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3901 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3961 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4021 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4081 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4141 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4201 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4261 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4321 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4381 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4441 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4501 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 4561 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 4621 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 4681 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 4741 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 4801 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 4861 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4921 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 5041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 5161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5461 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5701 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 5761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5881 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5941 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6001 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6061 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6121 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6181 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6241 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6301 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6361 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6421 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6481 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6541 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6601 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6661 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6721 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6781 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6841 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6901 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6961 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 7021 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat 7081 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 7141 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 7201 cttagg //