LOCUS V025936 8006 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025936 VERSION V025936 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..3496 /label="THSD1(NM_018676)" /note="THSD1(NM_018676)" /gene="THSD1" CDS 3503..3568 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3606..3830 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3876..4304 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4318..4647 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4714..5508 /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 5682..5803 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5852..5868) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5876..5892 /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(5900..5930) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5945..5966 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6254..6839) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7010..7870) /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(7871..7975) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgaaaccaa tgttgaaaga 961 cttttcaaat ctattgttgg tggtactctg tgactatgtt cttggagaag ctgaatatct 1021 tctcttgaga gagccaggcc atgtagcact aagcaacgac acagtgtatg tggatttcca 1081 gtattttgat ggtgctaatg ggacactgag gaatgtatct gtcctgctgt tggaggccaa 1141 caccaatcag actgtaacta ccaagtacct cctgaccaac cagtcccagg gaacactaaa 1201 gtttgagtgc ttctatttca aggaggctgg tgactactgg ttcacaatga ctccagaagc 1261 aacagacaac agcactccat tcccctggtg ggagaaaagt gcctttctga aggtggaatg 1321 gcctgtcttt cacgttgact tgaataggag tgccaaggca gcagaaggca ccttccaagt 1381 gggcctattt accagtcaac cactgtgccc gtttcctgtg gacaagccca acatcgtagt 1441 ggatgtcatc ttcaccaaca gtcttcctga ggcaagaaga aattcaagac agccgctgga 1501 aataagaacc agcaaaagga cagaacttgc tcaaggtcag tgggttgagt ttggctgtgc 1561 acccttgggg ccagaagcct atgtcaccgt ggtgctgaag ctgcttgggc gagactcagt 1621 cattacctcc acaggaccca ttgacctggc ccagaaattt ggatacaaac tggtgatggt 1681 gccagaactc acatgtgagt ccggggtaga ggtgacagtg ctgcctccac catgcacctt 1741 cgtccaagga gtggtcactg tcttcaagga ggcccccaga taccctggga agaggaccat 1801 tcacttggct gaaaacagcc tgcccctggg agagaggagg acaattttta actgtacttt 1861 gtttgacatg gggaagaata agtactgctt tgactttggc atttcaagca gaagccattt 1921 ttctgcaaag gaggagtgca tgctaattca gagaaataca gaaacttggg gactgtggca 1981 gccatggagc cagtgtagtg ccacatgtgg ggatggtgtc agagagcgtc gccgagtgtg 2041 tctcacttcc ttcccctcca gtcctgtctg ccctggaatg tccttggagg cctccctgtg 2101 ttccctggag gagtgtgctg ctttccagcc atccagccca tctcctcttc agccccaggg 2161 tccagtgaag tccaacaaca tcgtgactgt cactggtata tccttgtgct tgttcatcat 2221 cattgccact gtgctcatca cgctgtggag gaggttcggc cggccagcca agtgcagcac 2281 acctgctcga cacaactcca tccactcccc cagcttccgg aagaactcgg acgaggagaa 2341 tatctgcgag ctgagcgagc agcgcgggag cttctcggat gggggagacg ggcccacggg 2401 gagtccaggg gacacaggca tccctctgac ctacaggcgg agcgggccgg tacctcccga 2461 ggatgatgcc tctggcagcg agagcttcca gtccaacgcc cagaagataa tcccacctct 2521 gttcagctac cgccttgccc agcagcagtt aaaggagatg aaaaagaaag gtctgacgga 2581 aactaccaaa gtgtatcacg tgtctcagag tcccctgaca gacactgcca ttgatgcggc 2641 ccccagcgct cccttagatt tggaaagccc ggaagaagct gcagcaaaca agttccggat 2701 caaatcccca tttccggagc agcccgcggt cagtgccggg gaaaggcctc cctccaggct 2761 ggatctaaat gtgactcagg ccagttgtgc cataagcccc agccagactc tgatccgcaa 2821 gtcacaggca aggcacgtgg gcagcagagg gggcccgtcc gaaaggagcc atgccaggaa 2881 cgcccatttc aggaggacag cgagtttcca tgaagccagg caggcccggc cgttccgaga 2941 gaggagcatg tccactctga ctccacggca ggcccctgcc tacagctcta ggacgcggac 3001 ctgcgagcag gcagaggaca gatttaggcc tcagagtcga ggtgcccacc tgtttcctga 3061 aaaactggag catttccaag aggcaagtgg aacccgtggt ccattaaacc ctctccctaa 3121 atcctacact ttggggcagc ccttgaggaa accagacctt ggggatcacc aggcaggatt 3181 agtggccgga attgagagaa cagagcccca cagagctcgt cggggaccgt cccccagtca 3241 caagagtgtc tcaaggaagc agtcttctcc catatccccc aaagataact accagagggt 3301 cagttctctg agcccttctc agtgtagaaa agacaagtgt caaagcttcc ccactcaccc 3361 tgagtttgcc ttctatgaca atacgtcgtt tggcctcact gaggctgagc agaggatgct 3421 ggacctccca ggatattttg ggtcaaatga agaggatgaa accacaagta cacttagcgt 3481 ggagaagctg gtgatcctcg aggactacaa agaccatgac ggtgattata aagatcatga 3541 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 3601 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3661 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3721 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3781 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3841 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3901 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3961 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4021 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4081 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4141 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4201 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4261 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4321 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4381 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4441 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4501 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4561 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4621 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4681 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4741 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4801 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4861 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4921 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4981 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 5041 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 5101 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 5161 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5221 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5281 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5341 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5401 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5461 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5521 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5581 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5641 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5701 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5761 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5821 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5881 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5941 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 6001 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 6061 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 6121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 6181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 6241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6781 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 7021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 7081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 7141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 7981 ttccccgaaa agtgccacct gacgtc //