pMXB10

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载体基本信息

出品公司: NEB(New England Biolabs)
载体名称: pMXB10
质粒类型: 原核细胞表达载体
高拷贝/低拷贝: --
启动子: T7
克隆方法: 多克隆位点,限制性内切酶
载体大小: 7816bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: 氨苄青霉素(Ampicillin)
筛选标记: --
备注: --
产品目录号: E6903
稳定性: --
组成型: --
病毒/非病毒: --

载体质粒图谱和多克隆位点信息

pMXB10载体图谱



载体简介

pMXB10 is a control plasmid for the IMPACT Kit (NEB #E6901) (1,2). This plasmid carries the 
E. coli malE gene, encoding the maltose binding protein (MBP, 42 kDa) (3), fused in-frame to 
the coding region of the Mxe gyrase A intein-chitin binding domain (28 kDa) from pTXB1 (2,4,5).
pMXB10 can be used to test plasmid transformation, cell culture, induction and purification 
procedures. After induction with 0.3 mM IPTG at 30°C for 3 hours (or 15°C for 12-16 hours), 
100 ml of cells should yield 2-3 mg of a 70 kDa fusion protein. After chitin column purification
and cleavage, approximately 1.5-2.0 mg of MBP is usually obtained. This double stranded vector 
is 7813 bp in length.

载体序列

LOCUS       pMXB10                  7816 bp    DNA     circular     19-NOV-2010
DEFINITION  Control vector pMXB10, complete sequence.
ACCESSION
VERSION
KEYWORDS    .
SOURCE      Control vector pMXB10
  ORGANISM  Control vector pMXB10
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 7816)
  AUTHORS   Telenti,A., Southworth,M., Alcaide,F., Daugelat,S., Jacobs,W.R. Jr.
            and Perler,F.B.
  TITLE     The Mycobacterium xenopi GyrA protein splicing element:
            characterization of a minimal intein
  JOURNAL   J Bacteriol 179, 6378-6382 (1997)
   PUBMED   9335286
REFERENCE   2  (bases 1 to 7816)
  AUTHORS   Evans,T.C. Jr., Benner,J. and Xu,M.-Q.
  TITLE     Semisynthesis of cytotoxic proteins using a modified protein
            splicing element
  JOURNAL   Protein Sci 7, 2256-2264 (1998)
   PUBMED   9827992
REFERENCE   3  (bases 1 to 7816)
  AUTHORS   Southworth,M.W., Amaya,K., Evans,T.C., Xu,M.-Q. and Perler,F.B.
  TITLE     Purification of proteins fused to either the amino or carboxy
            terminus of the Mycobacterium xenopi Gyrase A intein
  JOURNAL   BioTechniques 27, 110-120 (1999)
   PUBMED   10407673
REFERENCE   4  (bases 1 to 7816)
  AUTHORS   Perler,F.B.
  TITLE     InBase, the intein database
  JOURNAL   Nucleic Acids Res 30 (1), 383-384 (2002)
   PUBMED   11752343
REFERENCE   5  (bases 1 to 7816)
  AUTHORS   Ghosh,I. and Xu,M.Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2007) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA
FEATURES             Location/Qualifiers
     source          1..7816
                     /organism="Control vector pMXB10"
                     /mol_type="other DNA"
     gene            140..1000
                     /gene="bla"
     CDS             140..1000
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"

     sig_peptide     140..208
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein"
     rep_origin      1042..1555
                     /note="M13 origin of replication (+ -)"
     rep_origin      1666..2254
                     /note="pMB1 origin of replication (clockwise) (RNAII -35
                     to RNA/DNA switch point)"
     gene            complement(2623..2814)
                     /gene="rop"
     CDS             complement(2623..2814)
                     /gene="rop"
                     /exception="alternative start codon"
                     /codon_start=1
                     /transl_table=11
                     /product="ROP protein"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
     gene            complement(3371..4453)
                     /gene="lacI"
     CDS             complement(3371..4453)
                     /gene="lacI"
                     /exception="alternative start codon"
                     /codon_start=1
                     /transl_table=11
                     /product="lac repressor (LacI)"

     primer_bind     5637..5656
                     /note="T7 universal primer sequence (NEB #S1248S)"
     promoter        5637..5654
                     /note="T7 promoter (transcript start 5654 clockwise)"
     protein_bind    5656..5680
                     /note="lac operator"
                     /bound_moiety="LacI"
     RBS             5710..5716
                     /note="RBS for expression CDS"
     gene            5725..7668
                     /gene="expression CDS"
     CDS             5725..7668
                     /gene="expression CDS"
                     /note="5' multiple cloning site (NdeI-NcoI) 5722..5740;
                     malE sequence 5743..6843; 3' multiple cloning site
                     6849..6882; Mxe GyrA intein sequence 6886..7479; chitin
                     binding domain (CBD) 7510..7668"
                     /codon_start=1
                     /product="MBP/intein/CBD fusion"

     primer_bind     complement(6936..6953)
                     /note="Mxe intein reverse primer sequence (NEB #S1268S,
                     discontinued; replaced by #S1285S)"
     primer_bind     complement(6996..7015)
                     /note="Mxe intein reverse II primer sequence (NEB
                     #S1285S)"
     terminator      7688..7810
                     /note="T7 Tphi transcription terminator"
BASE COUNT     1931 a   2021 c   2138 g   1726 t
ORIGIN
        1 aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
       61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      301 ccttgagagt tttcgccccg aagaacgttc tccaatgatg agcactttta aagttctgct
      361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
      961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1021 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag
     1081 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1201 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1321 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1381 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1441 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1501 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag
     1561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2401 cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca atctgctctg
     2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     2701 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     2761 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     2821 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     2881 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     2941 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcc
     3001 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     3061 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3121 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     3181 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     3241 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     3661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3841 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4501 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4561 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4621 aatggtgcat gccggcatgc cgccctttcg tcttcaagaa ttaattccca attccccagg
     4681 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     4741 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     4801 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc
     4861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4921 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4981 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc
     5041 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     5101 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5161 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa
     5221 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     5281 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     5341 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     5401 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5461 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     5521 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     5581 aattggggat cggaattaat tcccggttta aaccggggat ctcgatcccg cgaaattaat
     5641 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5701 taactttaag aaggagatat acatatggga agagccatgg atatgaaaac tgaagaaggt
     5761 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
     5821 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
     5881 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
     5941 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
     6001 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
     6061 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
     6121 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
     6181 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
     6241 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
     6301 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat
     6361 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
     6421 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
     6481 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
     6541 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg
     6601 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
     6661 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccactatgga aaacgcccag
     6721 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
     6781 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
     6841 actaattcga gctcgaagct tggcggccgc gaattcctcg agtactgcat cacgggagat
     6901 gcactagttg ccctacccga gggcgagtcg gtacgcatcg ccgacatcgt gccgggtgcg
     6961 cggcccaaca gtgacaacgc catcgacctg aaagtccttg accggcatgg caatcccgtg
     7021 ctcgccgacc ggctgttcca ctccggcgag catccggtgt acacggtgcg tacggtcgaa
     7081 ggtctgcgtg tgacgggcac cgcgaaccac ccgttgttgt gtttggtcga cgtcgccggg
     7141 gtgccgaccc tgctgtggaa gctgatcgac gaaatcaagc cgggcgatta cgcggtgatt
     7201 caacgcagcg cattcagcgt cgactgtgca ggttttgccc gcggaaaacc cgaatttgcg
     7261 cccacaacct acacagtcgg cgtccctgga ctggtgcgtt tcttggaagc acaccaccga
     7321 gacccggacg cccaagctat cgccgacgag ctgaccgacg ggcggttcta ctacgcgaaa
     7381 gtcgccagtg tcaccgacgc cggcgtgcag ccggtgtata gccttcgtgt cgacacggca
     7441 gaccacgcgt ttatcacgaa cgggttcgtc agccacgcta ctggcctgac cggtctgaac
     7501 tcaggcctca cgacaaatcc tggtgtatcc gcttggcagg tcaacacagc ttatactgcg
     7561 ggacaattgg tcacatataa cggcaagacg tataaatgtt tgcagcccca cacctccttg
     7621 gcaggatggg aaccatccaa cgttcctgcc ttgtggcagc ttcaatgact gcaggaaggg
     7681 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     7741 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     7801 ggaactatat ccggat

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