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
//
