pMal-c5x-NO09G02500.1

LOCUS       pMal-c5x-NO09G02500.1            7849 bp DNA     circular SYN 24-JUL-2024
DEFINITION  Bacterial vector for inducible cytoplasmic expression of 
            maltose-binding protein (MBP) fusions with a Factor Xa cleavage 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    pMAL-c5X
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7849)
  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 7849)
  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..7849
                     /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"
     primer_bind     complement(4873..4899)
                     /label=引物 1
     terminator      5113..5140
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5168..5258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5259..6119
                     /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      6207..6795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7165..7356)
                     /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 tccatgaggc agtcctccaa gcatgcctca
     2761 accatcctgc cccttctcct cctcgtccac ttcctggcag tctgcacgca tgccttccag
     2821 tcccttcgcg cactgaagtg tccgctcgtc acgcagctgt caaatgccca ttcctcttcg
     2881 tggtcctcca ctgacacgga aactcggcca gagcgagaga cagtgatcaa gtccgcctca
     2941 tcccaaccat cacaagcacc ccaaaccccc ccaaaacgcc agccctggcg ccgtctgggc
     3001 agctttgaga cattgttggc ccggaaagca gcagcaggac cgcagcggtt gtccatacct
     3061 catgtctgtg tggccgttgt cgatgatgaa caaggactgt tgaatgctcg gctgcttaag
     3121 ccagcgttag tctcgctcgt gcggcgccat cccaagctgc gagcctgcat tcagggcacg
     3181 ggcaaggtgg agggagcgcc tgtggccggg gtgcgacgag gaggaagaga agtcgacccc
     3241 ttgatgtggt ggccatgtgc gatgacacct gaagaggtgg tagaagaagc gttgtcattc
     3301 gacgacggca gcagtagcag cagcattaat gacgatgcgc aggactctcc tttcttcttc
     3361 tctgtatcct ttcaagcggc tttggacaat acggacatgg attcgagtcg tggacctctg
     3421 tggcgactgc atgtaatgcg ctctcaaggg agcagaaaag gcaccaaacc tgcagggcca
     3481 ggaggagtag ggcaggttgc tttgctcttc attttcaacc acgccataag cgaccaagtc
     3541 tcctgcaaca cgctcatcca tgagctactg tcgacgctcg gagagacctc atcctcatcc
     3601 tcatcctcat cctcatcctc ctcctccatc gcttcccagc ctttcccccc ctccatcgaa
     3661 aaggccatcc tgggtcccgc ccgagccacg gtcagtttca caactgccct ctacgccgcc
     3721 cgcgaatgtc ttttcggcct tctccccacc actctctacc ctacgcctac agcaacagca
     3781 gcagcgacag ggaccgcgac agcactagac accatccctc ctcccctccc tgcagcgaac
     3841 cgaaggacca tatgtgtatt tcgtcgtgag gaaccctcca ggctgtccct cttgaaggaa
     3901 gagtgtaaga agaggggggt gacagtgaca gctgccttgt ctgcggccat gctgtatgcc
     3961 accagtgatc tggcgcatgg agaggaagag gaagggaggg ctgaaggggt acgaaacgga
     4021 aaagagcaca attatcgttt tttgctgtcc ttgaacatgc gcacattttc tcgccgtaag
     4081 cttgggggag gaacggaggg aggggaggga gaaaaggaga aagggaagga tgacaaggag
     4141 ggggaggagg actgggaaga gaatgcagtg gcatcagcag ccggtgcaat ggactttatg
     4201 gtccgcttgc aggaaaggca aggcagaaac ctcctctgta ctcacaaatc aacgacaagc
     4261 aagaacggaa cggtagccgc aactgctgca acagcaccag gagcaccaag accacatgta
     4321 gctggagagg aagcattttg gctcctggct aaacaatgcc gtcgtgagtt tctagatttt
     4381 gtctcggccg gtttcgtccc cgaatccgtt aatttgtttg attggggtat gcagacattg
     4441 gaattgaatg aggtagtgga gaaggaggcc gaaaacccta agactctggg aagggcgtat
     4501 acttgtggag tgtcgaacat gggagtgttt gaagatggga tgggaggagg gaggcgcaga
     4561 ggcagcaatc gcagcagcaa cggcagccga gggatggacg aagatttctc tctctcttct
     4621 tcagcctttt cctcttgtct accctcctat ggatctctcc gactccacgg aattcactat
     4681 gcaaccagtc acagcttgac gggaagcctg taccagctgt cgtgcgggac ggtaaatggg
     4741 gcgctatgtc tgacattcca ctacccctgg ccgttggtgg atcgagagat ggccaatcgg
     4801 ttcgcggaca gagtggttca aatcttgagg tgtattggtt ggggagagag aacggaggga
     4861 gggagggagg gagggaggga cagagaagag gaaagttgag gatccgaatt ccctgcaggt
     4921 aattaaataa gcttcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     4981 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     5041 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     5101 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     5161 cggtccgttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5221 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5281 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5341 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5401 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttc ccaatgatga
     5461 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     5521 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5581 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5641 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5701 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5761 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5821 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5881 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5941 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6001 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6061 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6121 tgtcagacca agtttactca tatatacttt agattgattt ccttaggact gagcgtcaac
     6181 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6241 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6301 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6361 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6421 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6481 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6541 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6601 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6661 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6721 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6781 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6841 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6901 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6961 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     7021 tcacaccgca tataaggtgc actgtgactg ggtcatggct gcgccccgac acccgccaac
     7081 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     7141 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     7201 gcagctgcgg taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc
     7261 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     7321 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     7381 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     7441 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     7501 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     7561 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     7621 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     7681 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     7741 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     7801 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgaaatt
//