LOCUS pET-61-DEST. 6846 bp DNA circular SYN 01-JAN-1980 DEFINITION Gateway(R) Nova bacterial destination vector for expressing proteins tagged at the N-terminus with 6xHis and Strep-Tag II. ACCESSION . VERSION . KEYWORDS pET-61-DEST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6846) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission REFERENCE 2 (bases 1 to 6846) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6846 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 12..467 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(585..1442) /label=AmpR /note="beta-lactamase" promoter complement(1443..1535) /label=AmpR promoter rep_origin 1616..2204 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2390..2532) /label=bom /note="basis of mobility region from pBR322" CDS complement(2637..2825) /label=rop /note="Rop protein, which maintains plasmids at low copy number" protein_bind complement(3363..3384) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(3400..4479) /label=lacI /note="lac repressor" promoter complement(4480..4557) /label=lacI promoter promoter 4870..4888 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 4889..4913 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." RBS 4928..4950 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4957..4959 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 4963..4980 /label=6xHis /note="6xHis affinity tag" CDS 4987..5010 /label=Strep-Tag II /note="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" protein_bind 5020..5144 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 5169..5199 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 5253..5909 /label=CmR /note="chloramphenicol acetyltransferase" CDS 6254..6556 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" protein_bind complement(6600..6724) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" terminator 6774..6821 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat 481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg 1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac 1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt 2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac 2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg 2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg 2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga 2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg 2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa 2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt 2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt 3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct 3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac 3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag 3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc 3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc 3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc 3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt 3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg 3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc 3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat 3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt 3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc 3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc 3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag 3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt 4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat 4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag 4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac 4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg 4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg 4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt 4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc 4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact 4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg 4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga 4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc 4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt 4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg 4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc 4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag 4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac 4981 gcaagctgga gccacccgca gttcgaaaag ggtgcagtca caagtttgta caaaaaagct 5041 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa 5101 cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc gcattaggca 5161 ccccaggctt tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc 5221 cgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca 5281 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc 5341 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga 5401 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc 5461 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc 5521 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc 5581 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa 5641 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct 5701 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg 5761 ttttcactat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc 5821 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac 5881 agtactgcga tgagtggcag ggcggggcgt aaacgcgtgg atccggctta ctaaaagcca 5941 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt 6001 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga 6061 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc 6121 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag 6181 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac 6241 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct 6301 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct 6361 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat 6421 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tttccgttat 6481 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct 6541 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc 6601 atagtgactg gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct 6661 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag 6721 tggtgtaagt gattaattga gcagcctagg ctgctgccac cgctgagcaa taactagcat 6781 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat 6841 ccggat //