LOCUS Exported 5677 bp DNA circular SYN 13-SEP-2025 DEFINITION Bacterial vector for inducible cytoplasmic expression of maltose-binding protein (MBP) fusions with a Factor Xa cleavage site. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5677) AUTHORS Riggs P. TITLE Expression and purification of maltose-binding protein fusions. JOURNAL Curr Protoc Mol Biol 2001;Chapter 16:Unit16.6. PUBMED 18265133 REFERENCE 2 (bases 1 to 5677) AUTHORS New England Biolabs TITLE Direct Submission REFERENCE 3 (bases 1 to 5677) TITLE Direct Submission REFERENCE 4 (bases 1 to 5677) TITLE Direct Submission REFERENCE 5 (bases 1 to 5677) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 13, 2025 from SnapGene 6.0.2 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr Protoc Mol Biol"; date: "2001"; volume: "Chapter 16"; pages: "Unit16.6" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..5677 /mol_type="other DNA" /organism="synthetic DNA construct" source join(4349..5677,1..4348) /mol_type="other DNA" /organism="synthetic DNA construct" promoter 77..105 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 113..129 /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)." CDS 199..1299 /codon_start=1 /label=MBP /note="maltose binding protein from E. coli" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" CDS 1348..1359 /codon_start=1 /label=Factor Xa site /note="Factor Xa recognition and cleavage site" /translation="IEGR" terminator 1434..1520 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1612..1639 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1666..1757 /label=AmpR promoter CDS 1758..2615 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2706..3294 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3667..3855) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" promoter 4351..4428 /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 4429..5508 /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" protein_bind 5524..5545 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat 61 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc 121 ggataacaat ttcacacagg aaacagccag tccgtttagg tgttttcacg agcaattgac 181 caacaaggac catagattat gaaaatcgaa gaaggtaaac tggtaatctg gattaacggc 241 gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt 301 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact 361 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct 421 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc 481 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta 541 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg 601 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa 661 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac 721 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc 781 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca 841 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc 901 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa 961 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa 1021 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt 1081 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggtgaaa 1141 gatccgcgta ttgccgccac tatggaaaac gcccagaaag gtgaaatcat gccgaacatc 1201 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt 1261 cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc gaacaacaac 1321 aacaataaca ataacaacaa cctcgggatc gagggaagga tttcacatat gtccatgggc 1381 ggccgcgata tcgtcgacgg atccgaattc cctgcaggta attaaataag cttcaaataa 1441 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg 1501 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg 1561 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca 1621 tcctgacgga tggccttttt gcgtttctac aaactctttc ggtccgttgt ttatttttct 1681 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 1741 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 1801 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 1861 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 1921 ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa gttctgctat 1981 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact 2041 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 2101 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 2161 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 2221 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 2281 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 2341 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 2401 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 2461 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 2521 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 2581 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 2641 atatacttta gattgatttc cttaggactg agcgtcaacc ccgtagaaaa gatcaaagga 2701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 2761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 2821 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 2881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 2941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 3001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 3061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 3121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 3181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 3241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 3301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 3361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 3421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 3481 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat ataaggtgca 3541 ctgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg 3601 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 3661 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc 3721 agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag 3781 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt 3841 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat 3901 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt 3961 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat 4021 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca 4081 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc 4141 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt 4201 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt 4261 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg 4321 tggccaggac ccaacgctgc ccgaaattcc gacaccatcg aatggtgcaa aacctttcgc 4381 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta 4441 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg 4501 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag 4561 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt 4621 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa 4681 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc 4741 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt 4801 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg 4861 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa 4921 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg 4981 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat 5041 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc 5101 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt 5161 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt 5221 ggtgcggata tttcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg 5281 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg 5341 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg 5401 aaaagaaaaa ccaccctggc gcccaataca caaaccgcct ctccccgcgc gttggccgat 5461 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 5521 aattaatgta agttagctca ctcattaggc acaattctca tgtttgacag cttatcatcg 5581 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg 5641 tgcaggtcgt aaatcactgc ataattcgtg tcgctca //