LOCUS V018695 8246 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018695 VERSION V018695 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 932..3721 /label="ADAMTS5(NM_007038)" /note="ADAMTS5(NM_007038)" /gene="ADAMTS5" CDS 3743..3808 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3846..4070 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4116..4544 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4558..4887 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4954..5748 /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 5922..6043 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6092..6108) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6116..6132 /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(6140..6170) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6185..6206 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6494..7079) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7250..8110) /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(8111..8215) /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 gctggctagt 901 taagcttggt accgagctcg gatccgccac catgctgctc gggtgggcgt ccctgctgct 961 gtgcgcgttc cgcctgcccc tggccgcggt cggccccgcc gcgacacctg cccaggataa 1021 agccgggcag cctccgactg ctgcagcagc cgcccagccc cgccggcggc agggggagga 1081 ggtgcaggag cgagccgagc ctcccggcca cccgcacccc ctggcgcagc ggcgcaggag 1141 caaggggctg gtgcagaaca tcgaccaact ctactccggc ggcggcaagg tgggctacct 1201 cgtctacgcg ggcggccgga ggttcctctt ggacctggag cgagatggtt cggtgggcat 1261 tgctggcttc gtgcccgcag gaggcgggac gagtgcgccc tggcgccacc ggagccactg 1321 cttctatcgg ggcacagtgg acggtagtcc ccgctctctg gctgtctttg acctctgtgg 1381 gggtctcgac ggcttcttcg cggtcaagca cgcgcgctac accctaaagc cactgctgcg 1441 cggaccctgg gcggaggaag aaaaggggcg cgtgtacggg gatgggtccg cacggatcct 1501 gcacgtctac acccgcgagg gcttcagctt cgaggccctg ccgccgcgcg ccagctgcga 1561 aacccccgcg tccacaccgg aggcccacga gcatgctccg gcgcacagca acccgagcgg 1621 acgcgcagca ctggcctcgc agctcttgga ccagtccgct ctctcgcccg ctgggggctc 1681 aggaccgcag acgtggtggc ggcggcggcg ccgctccatc tcccgggccc gccaggtgga 1741 gctgcttctg gtggctgacg cgtccatggc gcggttgtat ggccggggcc tgcagcatta 1801 cctgctgacc ctggcctcca tcgccaatag gctgtacagc catgctagca tcgagaacca 1861 catccgcctg gccgtggtga aggtggtggt gctaggcgac aaggacaaga gcctggaagt 1921 gagcaagaac gctgccacca cactcaagaa cttttgcaag tggcagcacc aacacaacca 1981 gctgggagat gaccatgagg agcactacga tgcagctatc ctgtttactc gggaggattt 2041 atgtgggcat cattcatgtg acaccctggg aatggcagac gttgggacca tatgttctcc 2101 agagcgcagc tgtgctgtga ttgaagacga tggcctccac gcagccttca ctgtggctca 2161 cgaaatcgga catttacttg gcctctccca tgacgattcc aaattctgtg aagagacctt 2221 tggttccaca gaagataagc gcttaatgtc ttccatcctt accagcattg atgcatctaa 2281 gccctggtcc aaatgcactt cagccaccat cacagaattc ctggatgatg gccatggtaa 2341 ctgtttgctg gacctaccac gaaagcagat cctgggcccc gaagaactcc caggacagac 2401 ctacgatgcc acccagcagt gcaacctgac attcgggcct gagtactccg tgtgtcccgg 2461 catggatgtc tgtgctcgcc tgtggtgtgc tgtggtacgc cagggccaga tggtctgtct 2521 gaccaagaag ctgcctgcgg tggaagggac gccttgtgga aaggggagaa tctgcctgca 2581 gggcaaatgt gtggacaaaa ccaagaaaaa atattattca acgtcaagcc atggcaactg 2641 gggatcttgg ggatcctggg gccagtgttc tcgctcatgt ggaggaggag tgcagtttgc 2701 ctatcgtcac tgtaataacc ctgctcccag aaacaacgga cgctactgca cagggaagag 2761 ggccatctac cgctcctgca gtctcatgcc ctgcccaccc aatggtaaat catttcgtca 2821 tgaacagtgt gaggccaaaa atggctatca gtctgatgca aaaggagtca aaacttttgt 2881 ggaatgggtt cccaaatatg caggtgtcct gccagcggat gtgtgcaagc tgacctgcag 2941 agccaagggc actggctact atgtggtatt ttctccaaag gtgaccgatg gcactgaatg 3001 taggctgtac agtaattccg tctgcgtccg ggggaagtgt gtgagaactg gctgtgacgg 3061 catcattggc tcaaagctgc agtatgacaa gtgcggagta tgtggaggag acaactccag 3121 ctgtacaaag attgttggaa cctttaataa gaaaagtaag ggttacactg acgtggtgag 3181 gattcctgaa ggggcaaccc acataaaagt tcgacagttc aaagccaaag accagactag 3241 attcactgcc tatttagccc tgaaaaagaa aaacggtgag taccttatca atggaaagta 3301 catgatctcc acttcagaga ctatcattga catcaatgga acagtcatga actatagcgg 3361 ttggagccac agggatgact tcctgcatgg catgggctac tctgccacga aggaaattct 3421 aatagtgcag attcttgcaa cagaccccac taaaccatta gatgtccgtt atagcttttt 3481 tgttcccaag aagtccactc caaaagtaaa ctctgtcact agtcatggca gcaataaagt 3541 gggatcacac acttcgcagc cgcagtgggt cacgggccca tggctcgcct gctctaggac 3601 ctgtgacaca ggttggcaca ccagaacggt gcagtgccag gatggaaacc ggaagttagc 3661 aaaaggatgt cctctctccc aaaggccttc tgcgtttaag caatgcttgt tgaagaaatg 3721 tctcgagtct agagggccct tcgactacaa agaccatgac ggtgattata aagatcatga 3781 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 3841 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3901 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3961 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 4021 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 4081 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 4141 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 4201 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4261 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4321 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4381 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4441 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4501 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4561 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4621 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4681 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4741 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4801 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4861 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4921 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4981 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 5041 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 5101 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 5161 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 5221 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 5281 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 5341 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 5401 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5461 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5521 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5581 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5641 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5701 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5761 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5821 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5881 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5941 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 6001 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 6061 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 6121 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 6181 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 6241 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 6301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 6361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 6421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 7021 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 7081 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 7141 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 7201 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 7261 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 7321 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 7381 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7441 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7501 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7561 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7621 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7681 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7741 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7801 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7861 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7921 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7981 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 8041 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 8101 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 8161 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 8221 ttccccgaaa agtgccacct gacgtc //