pMAL-p5x

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载体基本信息

出品公司: NEB
载体名称: pMAL-p5x
质粒类型: 大肠杆菌表达载体;蛋白表达与纯化
高拷贝/低拷贝: 低拷贝
启动子: tac
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5752bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: MBP tag
载体抗性: Ampicillin
筛选标记: --
备注: --
产品目录号: N8109S
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

pMAL-p5x载体图谱



pMAL-p5x多克隆位点

pMAL-p5x载体特征

载体简介


pMAL-p5X-His Vector

The vector pMAL-p5X is designed to produce maltose-binding protein (MBP) fusions, 
where the protein of interest can be cleaved from MBP with the specific protease Factor Xa.

MBP fusions made with this vector include an N-terminal signal sequence, so the 
fusion protein is directed to the periplasm. The MBP has been engineered for tighter 
binding to amylose resin.

A gene or open reading frame is inserted into a restriction site of the vector polylinker, 
in the same translational reading frame as the malE gene (encoding maltose-binding protein).
The fusion protein thus produced can be purified by amylose affinity chromatography. The 
sequence coding for the four amino acids Ile-Glu-Gly-Arg is present just upstream of the XmnI 
site. This allows the protein of interest to be cleaved from maltose-binding protein with 
the specific protease Factor Xa. Fragments inserted in the XmnI site (cleaves GAAGG↓ATTTC) 
will produce a fusion protein that, after Factor Xa cleavage, contains no vector-derived 
residues on the protein of interest.

载体序列

LOCUS       pMAL-p5X                5752 bp    DNA     circular     10-NOV-2008
DEFINITION  Cloning vector pMAL-p5X, complete sequence.
ACCESSION
VERSION
KEYWORDS    .
SOURCE      Cloning vector pMAL-p5X
  ORGANISM  Cloning vector pMAL-p5X
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 5752)
  AUTHORS   Riggs,P.
  TITLE     Expression and purification of maltose-binding protein fusions
  JOURNAL   (in) Asubel,F.A., Brent,R., Kingston,R.E., Moore,D.D.,
            Seidman,J.G., Smith,J.A. and Struhl,K. (Eds.);
            CURRENT PROTOCOLS IN MOLECULAR BIOLOGY: 1-14;
            Greene Publishing and Wiley Interscience, New York, NY (1992)
REFERENCE   2  (bases 1 to 5752)
  AUTHORS   Riggs,P.D.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-NOV-2008) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA
FEATURES             Location/Qualifiers
     source          1..5752
                     /organism="Cloning vector pMAL-p5X"
                     /mol_type="other DNA"
     gene            15..1163
                     /gene="lacIq"
     -35_signal      15..19
                     /gene="lacIq"
                     /note="lacIq promoter (clockwise); TGGTG"
     -10_signal      40..46
                     /gene="lacIq"
                     /note="lacIq promoter (clockwise); CATGATA"
     CDS             81..1163
                     /gene="lacIq"
                     /codon_start=1
                     /transl_table=11
                     /product="lac repressor (LacI)"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
     gene            1406..3043
                     /gene="malE expression cassette"
     -35_signal      1406..1411
                     /gene="malE expression cassette"
                     /note="Ptac promoter (clockwise); TTGACA"
     -10_signal      1428..1433
                     /gene="malE expression cassette"
                     /note="Ptac promoter (clockwise); TATAAT"
     CDS             1528..2829
                     /gene="malE expression cassette"
                     /note="malE sequence 1528..2703; spacer and factor Xa site
                     2704..2762; multiple cloning site region 2763..2829"
                     /codon_start=1
                     /product="maltose binding protein (MBP) fusion"
                     /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN
                     GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL
                     AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA
                     LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL
                     TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF
                     KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE
                     EELVKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT
                     NSSSNNNNNNNNNNLGIEGRISHMSMGGRDIVDGSEFPAGN"
     primer_bind     2662..2685
                     /note="malE primer sequence (NEB #S1237S)"
     misc_feature    2770..2822
                     /gene="malE expression cassette"
                     /note="multiple cloning site (NdeI-SbfI)"
     terminator      2841..2884
                     /gene="malE expression cassette"
                     /note="rrnB T1 transcription terminator"
     primer_bind     complement(2898..2921)
                     /note="MCS reverse primer sequence (NEB #SxxxxS)"
     terminator      3016..3043
                     /gene="malE expression cassette"
                     /note="rrnB T2 transcription terminator"
     gene            3092..4022
                     /gene="bla"
     -35_signal      3092..3097
                     /gene="bla"
                     /note="bla promoter (clockwise); TTCAAA"
     -10_signal      3115..3120
                     /gene="bla"
                     /note="bla promoter (clockwise); GACAAT"
     CDS             3162..4022
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRHIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      4110..4698
                     /note="pMB1 origin of replication (clockwise) (RNAII -35
                     to RNA/DNA switch point)"
     gene            complement(5068..5259)
                     /gene="rop"
     CDS             complement(5068..5259)
                     /gene="rop"
                     /codon_start=1
                     /transl_table=11
                     /product="ROP protein"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
BASE COUNT     1439 a   1453 c   1536 g   1324 t
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 aaaataaaaa caggtgcacg catcctcgca
     1561 ttatccgcat taacgacgat gatgttttcc gcctcggctc tcgccaaaat cgaagaaggt
     1621 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
     1681 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
     1741 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
     1801 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
     1861 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
     1921 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
     1981 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
     2041 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
     2101 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
     2161 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat
     2221 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
     2281 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
     2341 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
     2401 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg
     2461 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
     2521 tcttacgagg aagagttggt gaaagatccg cgtattgccg ccactatgga aaacgcccag
     2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
     2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
     2701 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg gatcgaggga
     2761 aggatttcac atatgtccat gggcggccgc gatatcgtcg acggatccga attccctgca
     2821 ggtaattaaa taagcttcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2881 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     2941 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     3001 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     3061 tttcggtccg ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3121 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3181 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3241 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3301 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt ttcccaatga
     3361 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     3421 agcaactcgg tcgccacata cactattctc agaatgactt ggttgagtac tcaccagtca
     3481 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3541 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3601 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3661 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3721 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3781 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3841 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3901 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3961 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4021 aactgtcaga ccaagtttac tcatatatac tttagattga tttccttagg actgagcgtc
     4081 aaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4741 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4801 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4861 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4921 atttcacacc gcatataagg tgcactgtga ctgggtcatg gctgcgcccc gacacccgcc
     4981 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5041 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5101 gaggcagctg cggtaaagct catcagcgtg gtcgtgcagc gattcacaga tgtctgcctg
     5161 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5221 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     5281 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     5341 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     5401 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     5461 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     5521 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5581 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5641 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     5701 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaaa tt
//

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