LOCUS V029657 8185 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029657 VERSION V029657 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 964..3675 /label="TLR3(NM_003265)" /note="TLR3(NM_003265)" /gene="TLR3" CDS 3682..3747 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3785..4009 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4055..4483 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4497..4826 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4893..5687 /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 5861..5982 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6031..6047) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6055..6071 /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(6079..6109) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6124..6145 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6433..7018) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7189..8049) /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(8050..8154) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgagac agactttgcc ttgtatctac ttttgggggg gccttttgcc ctttgggatg 1021 ctgtgtgcat cctccaccac caagtgcact gttagccatg aagttgctga ctgcagccac 1081 ctgaagttga ctcaggtacc cgatgatcta cccacaaaca taacagtgtt gaaccttacc 1141 cataatcaac tcagaagatt accagccgcc aacttcacaa ggtatagcca gctaactagc 1201 ttggatgtag gatttaacac catctcaaaa ctggagccag aattgtgcca gaaacttccc 1261 atgttaaaag ttttgaacct ccagcacaat gagctatctc aactttctga taaaaccttt 1321 gccttctgca cgaatttgac tgaactccat ctcatgtcca actcaatcca gaaaattaaa 1381 aataatccct ttgtcaagca gaagaattta atcacattag atctgtctca taatggcttg 1441 tcatctacaa aattaggaac tcaggttcag ctggaaaatc tccaagagct tctattatca 1501 aacaataaaa ttcaagcgct aaaaagtgaa gaactggata tctttgccaa ttcatcttta 1561 aaaaaattag agttgtcatc gaatcaaatt aaagagtttt ctccagggtg ttttcacgca 1621 attggaagat tatttggcct ctttctgaac aatgtccagc tgggtcccag ccttacagag 1681 aagctatgtt tggaattagc aaacacaagc attcggaatc tgtctctgag taacagccag 1741 ctgtccacca ccagcaatac aactttcttg ggactaaagt ggacaaatct cactatgctc 1801 gatctttcct acaacaactt aaatgtggtt ggtaacgatt cctttgcttg gcttccacaa 1861 ctagaatatt tcttcctaga gtataataat atacagcatt tgttttctca ctctttgcac 1921 gggcttttca atgtgaggta cctgaatttg aaacggtctt ttactaaaca aagtatttcc 1981 cttgcctcac tccccaagat tgatgatttt tcttttcagt ggctaaaatg tttggagcac 2041 cttaacatgg aagataatga tattccaggc ataaaaagca atatgttcac aggattgata 2101 aacctgaaat acttaagtct atccaactcc tttacaagtt tgcgaacttt gacaaatgaa 2161 acatttgtat cacttgctca ttctccctta cacatactca acctaaccaa gaataaaatc 2221 tcaaaaatag agagtgatgc tttctcttgg ttgggccacc tagaagtact tgacctgggc 2281 cttaatgaaa ttgggcaaga actcacaggc caggaatgga gaggtctaga aaatattttc 2341 gaaatctatc tttcctacaa caagtacctg cagctgacta ggaactcctt tgccttggtc 2401 ccaagccttc aacgactgat gctccgaagg gtggccctta aaaatgtgga tagctctcct 2461 tcaccattcc agcctcttcg taacttgacc attctggatc taagcaacaa caacatagcc 2521 aacataaatg atgacatgtt ggagggtctt gagaaactag aaattctcga tttgcagcat 2581 aacaacttag cacggctctg gaaacacgca aaccctggtg gtcccattta tttcctaaag 2641 ggtctgtctc acctccacat ccttaacttg gagtccaacg gctttgacga gatcccagtt 2701 gaggtcttca aggatttatt tgaactaaag atcatcgatt taggattgaa taatttaaac 2761 acacttccag catctgtctt taataatcag gtgtctctaa agtcattgaa ccttcagaag 2821 aatctcataa catccgttga gaagaaggtt ttcgggccag ctttcaggaa cctgactgag 2881 ttagatatgc gctttaatcc ctttgattgc acgtgtgaaa gtattgcctg gtttgttaat 2941 tggattaacg agacccatac caacatccct gagctgtcaa gccactacct ttgcaacact 3001 ccacctcact atcatgggtt cccagtgaga ctttttgata catcatcttg caaagacagt 3061 gccccctttg aactcttttt catgatcaat accagtatcc tgttgatttt tatctttatt 3121 gtacttctca tccactttga gggctggagg atatcttttt attggaatgt ttcagtacat 3181 cgagttcttg gtttcaaaga aatagacaga cagacagaac agtttgaata tgcagcatat 3241 ataattcatg cctataaaga taaggattgg gtctgggaac atttctcttc aatggaaaag 3301 gaagaccaat ctctcaaatt ttgtctggaa gaaagggact ttgaggcggg tgtttttgaa 3361 ctagaagcaa ttgttaacag catcaaaaga agcagaaaaa ttatttttgt tataacacac 3421 catctattaa aagacccatt atgcaaaaga ttcaaggtac atcatgcagt tcaacaagct 3481 attgaacaaa atctggattc cattatattg gttttccttg aggagattcc agattataaa 3541 ctgaaccatg cactctgttt gcgaagagga atgtttaaat ctcactgcat cttgaactgg 3601 ccagttcaga aagaacggat aggtgccttt cgtcataaat tgcaagtagc acttggatcc 3661 aaaaactctg tacatctcga ggactacaaa gaccatgacg gtgattataa agatcatgac 3721 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc 3781 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 3841 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 3901 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 3961 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 4021 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 4081 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 4141 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 4201 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 4261 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 4321 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 4381 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 4441 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 4501 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 4561 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 4621 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 4681 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 4741 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 4801 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 4861 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 4921 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 4981 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 5041 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 5101 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 5161 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 5221 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 5281 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 5341 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 5401 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 5461 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 5521 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 5581 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 5641 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 5701 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 5761 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 5821 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 5881 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5941 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 6001 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 6061 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6121 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6181 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6241 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6301 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6361 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6421 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6481 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6541 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6601 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6661 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6721 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6781 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6841 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 6901 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6961 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7681 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7741 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7801 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7861 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7921 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7981 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8041 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8161 tccccgaaaa gtgccacctg acgtc //