pMal-c5x-Phatr3 J36930

LOCUS       pMal-c5x-Phatr3 J36930            8746 bp DNA     circular SYN 04-AUG-2026
DEFINITION  Bacterial vector for inducible cytoplasmic expression of 
            maltose-binding protein (MBP) fusions with a Factor Xa cleavage 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    2941-pMAL-c5X-Phatr3 J36930
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8746)
  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 8746)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Protoc
            Mol Biol"; date: "2001"; volume: "Chapter 16"; pages: "Unit16.6"
FEATURES             Location/Qualifiers
     source          1..8746
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..80
                     /gene="lacI (mutant)"
                     /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             81..1163
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    1406..1434
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1528..2628
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             2677..2688
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2734..4653
                     /label=1
     primer_bind     complement(3170..3204)
                     /label=引物 2
     misc_feature    3190..3193
                     /label=A
     primer_bind     3194..3219
                     /label=引物 1
     misc_feature    4654..5793
                     /label=2
     terminator      6010..6037
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6065..6155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6156..7016
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7104..7692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8062..8253)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatttcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcaattgacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcacatatg
     2701 tccatgggcg gccgcgatat cgtcgacgga tccatgatgc tccaatcgga taggatttca
     2761 cccattggcg tcgcgttact cctatacctg gcaacttcca ttggacacgc ctttgcacct
     2821 cgatcggtgc tgactttgcg ccgcattact cagccttcca acccacgcca agccagttcc
     2881 acggatgctg acgacggtgg catgttcttt gcaagtgcgt ccgatgaaac tttgctggct
     2941 cggcccggta atgccacgaa cgatgatgcc gctaccgctg atgcaggaca agtcaacaac
     3001 caatcctctc ggcaaagtcg ggttctggga tcacaggaac tactcatgct accacgtcaa
     3061 tacggtccca aaagggacaa ggtaatcttt ccacaaatga accacgtttc ctgtgcgatt
     3121 ctatcggcga cgccctccga caccgtcctg caaaaagcac tagatgctgc catgcaggct
     3181 caccccttgt tgcactgccg agtagatgga gacggcgagc ctgataaacg aatagatcta
     3241 tttcaaatgg ttcggcaagg cgaacccaat ccttgtacct ttgtgtcgga gcccaacttg
     3301 ttttcggccc tagacgtact caaggtcatc gacgtgccta gtaccgaccg ggaggccctc
     3361 gatcaatcat ggcagaaaag tttcaacaaa aacttggacg atgggagttg gtgtgacgcc
     3421 gcgaaaggcc cattgtggaa agtagaactc catagaactg catctagcag tggtccgtgc
     3481 gctttactat tctccttcaa ccacgccatt tccgatcaaa gcagtgccaa tcgccttctg
     3541 gatcaaattt tgcgtaacgt ggccgaactc gaaagcttag gatcaactac accggcgatt
     3601 ccgcagacca tgccgttggc gttagaagac tcggtcatgg gagtgtcgcg gcgctggagt
     3661 gacgtgcaga tggacggagt tggtaccgac accatccggt acgtagtcaa taaggccgcc
     3721 gaaggtttgc gcaatcctgt tatcgtaccg gattcggccg gacgaaaaga agggagtaat
     3781 ccactgaatg cccttaccat catttcaggc cgtgcggccg gcggtcaaga taacgaaagt
     3841 caatctcggc gatccacttt acaatttaga tcactctcgc gggaaagcac gtcagctttg
     3901 ctagccaagt gccgagaaaa tggagtgtcg ataagcaacg ccctgacggc agctgcggcc
     3961 ctcacggcga cagatgtcat cgacaatggt aagccaaaaa gaaaatcacg caactacaaa
     4021 gtattgcaat ccttggacat gagaagatac ggagcgaaac ttgaccaagg agaaagtgtc
     4081 gcttgcatgg cgggtagtat ggatttgatg cttgggcctc ttgcggatcg gtctggaagg
     4141 accctgaaac agacttatac gccggaagca gccactattt tttggaattt agcaagagaa
     4201 agtagaaagc aaacaatcga ctttatggag tcaaacggtc cagagcaagc agttagggtc
     4261 tttgattttg ctatgtccat ttctgatctc aacaatttag tccacttgac tgccgagtca
     4321 aaagacaata aaggacgtgc ctacagtgca ggtatcacca atgtgggtgt ctacgaaagt
     4381 caagaagctt ttgctcgcat tgatcagtct gagcccgctg ttctccttgt tgaacacgga
     4441 aagtacaaaa ttcaagatat attctttgca acaactcata cccgatcagg atgcctatac
     4501 ccagcttcct gtattactgt tgacggctcg ctcaaaatga cctttaatcc ggtagctcct
     4561 atcgtcagcg ataagactaa cgccaacttt gcggatgggt ttgtggaact tctcgagatt
     4621 gttgccgggg taagccaagg cacaattaaa aaggatgaag agaggatggg gctcttagcg
     4681 tccatcccag agaatgcgct ttcagctgca acggcagttg ttggactagc agcagtcttg
     4741 tcacacgccc cagcatggct agactttttt tcatccgtga acgaaatgaa gcagaacatt
     4801 caagacccgg ccgacttttg ggccgccctc aacttttgga tcttttttgc agttggacat
     4861 cctattctcc agcctattct gtggataagc gatgttctac acgcaagtcc tggtcctaag
     4921 atcggaggcc tcattccagc cacctttttg gccggaaacc ttgcatttat tgctttgatc
     4981 accaagtcca aagagattcg caatgcagtc aacattgccg cactcgcggc gttcttaacg
     5041 tacgttggct ctggcttgga tggaacggct ggcacaggcg actttaattt ggcactagat
     5101 gatagctaca aagggcaaat agtaagagga tgtccctctt atgacgaagt ccgtcagcca
     5161 agtatggaca attttgatct ggaaaagtat cagggcctat ggtatgagca aaaatttcat
     5221 gattggactc aattcaaaga agtatacgat acgacccttg atataaagct aaccgaagga
     5281 gggaagggtt gggttgatga ctttgctgtc aaagggccgg ctccagacgc tgccaagctc
     5341 tcgtgggaca agagccctgt tgccaacgga gctcactact ttttgtttgg acgagtggat
     5401 ccgaatgatc cgccaggtat tctacgggag aaaggcttcg gcgtcgaatt tccgaactac
     5461 attgtcgacg tgaagaaaga tcctcaaact ggcgattaca aggaagccat tcagtttcaa
     5521 tgcttggaac ggggtggcgt ccgcgtgttt gagggtatta atttcatgtc tcgcaatcct
     5581 accatgacag atgaggagat gactgcaatg cattcacgtg cagagcgagc gggtatgtac
     5641 ccctatggag ctagtccgga gcagatgcac actgtagcac gacgaccggt gaatgccccg
     5701 gagatcgaca acaattggca agccatgtgg cgtgctctag gcgttgacaa gctgctagaa
     5761 ctcttgacac aatcaatcga agatggaggg cgataaggat ccgaattccc tgcaggtaat
     5821 taaataagct tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     5881 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     5941 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     6001 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttcgg
     6061 tccgttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6121 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6181 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6241 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6301 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgtttccca atgatgagca
     6361 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     6421 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6481 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6541 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6601 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6661 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6721 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6781 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6841 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6901 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6961 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7021 cagaccaagt ttactcatat atactttaga ttgatttcct taggactgag cgtcaacccc
     7081 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7141 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7201 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     7261 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7321 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7381 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7441 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7501 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7561 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7621 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7681 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7741 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7801 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7861 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     7921 caccgcatat aaggtgcact gtgactgggt catggctgcg ccccgacacc cgccaacacc
     7981 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     8041 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     8101 gctgcggtaa agctcatcag cgtggtcgtg cagcgattca cagatgtctg cctgttcatc
     8161 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     8221 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     8281 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     8341 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     8401 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     8461 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     8521 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     8581 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     8641 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     8701 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gaaatt
//