443143416pMAL-p5E
- 產(chǎn)品貨號GS-2057
- 銷售價(jià)格￥ 680
- 包裝規(guī)格2μg質(zhì)粒或0.5ml甘油菌
- 表達(dá)類型高
- 載體抗性Ampicillin (氨芐青霉素)
- 載體大小5755 bp
- 啟動(dòng)子Tac
The vector pMAL-p5E is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease Enterokinase (NEB #P8070).
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 tigher 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 MBP). The fusion protein produced from the vector can be purified by amylose affinity chromatography. The sequence coding for the five amino acids Asp-Asp-Asp-Asp-Lys is present just upstream of the KpnI site. This allows the protein of interest to be cleaved from maltose-binding protein with enterokinase.
pMAL-p5E cut with KpnI followed by treatment with the Quick Blunting Kit (NEB #E1201) produces a blunt end at the lysine codon. This allows blunt-end cloning of an insert where the first three nucleotides code for the first amino acid of the protein of interest, and enterokinase cleavage of the fusion produces a protein with no vector-derived amino acids.
0001 CCGACACCAT CGAATGGTGC AAAACCTTTC GCGGTATGGC ATGATAGCGC CCGGAAGAGA
0061 GTCAATTCAG GGTGGTGAAT GTGAAACCAG TAACGTTATA CGATGTCGCA GAGTATGCCG
0121 GTGTCTCTTA TCAGACCGTT TCCCGCGTGG TGAACCAGGC CAGCCACGTT TCTGCGAAAA
0181 CGCGGGAAAA AGTGGAAGCG GCGATGGCGG AGCTGAATTA CATTCCCAAC CGCGTGGCAC
0241 AACAACTGGC GGGCAAACAG TCGTTGCTGA TTGGCGTTGC CACCTCCAGT CTGGCCCTGC
0301 ACGCGCCGTC GCAAATTGTC GCGGCGATTA AATCTCGCGC CGATCAACTG GGTGCCAGCG
0361 TGGTGGTGTC GATGGTAGAA CGAAGCGGCG TCGAAGCCTG TAAAGCGGCG GTGCACAATC
0421 TTCTCGCGCA ACGCGTCAGT GGGCTGATCA TTAACTATCC GCTGGATGAC CAGGATGCCA
0481 TTGCTGTGGA AGCTGCCTGC ACTAATGTTC CGGCGTTATT TCTTGATGTC TCTGACCAGA
0541 CACCCATCAA CAGTATTATT TTCTCCCATG AAGACGGTAC GCGACTGGGC GTGGAGCATC
0601 TGGTCGCATT GGGTCACCAG CAAATCGCGC TGTTAGCGGG CCCATTAAGT TCTGTCTCGG
0661 CGCGTCTGCG TCTGGCTGGC TGGCATAAAT ATCTCACTCG CAATCAAATT CAGCCGATAG
0721 CGGAACGGGA AGGCGACTGG AGTGCCATGT CCGGTTTTCA ACAAACCATG CAAATGCTGA
0781 ATGAGGGCAT CGTTCCCACT GCGATGCTGG TTGCCAACGA TCAGATGGCG CTGGGCGCAA
0841 TGCGCGCCAT TACCGAGTCC GGGCTGCGCG TTGGTGCGGA TATTTCGGTA GTGGGATACG
0901 ACGATACCGA AGACAGCTCA TGTTATATCC CGCCGTTAAC CACCATCAAA CAGGATTTTC
0961 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 GGATGACGAT
2761 GACAAGGTAC CGCATATGTC CATGGGCGGC CGCGATATCG TCGACGGATC CGAATTCCCT
2821 GCAGGTAATT AAATAAGCTT CAAATAAAAC GAAAGGCTCA GTCGAAAGAC TGGGCCTTTC
2881 GTTTTATCTG TTGTTTGTCG GTGAACGCTC TCCTGAGTAG GACAAATCCG CCGGGAGCGG
2941 ATTTGAACGT TGCGAAGCAA CGGCCCGGAG GGTGGCGGGC AGGACGCCCG CCATAAACTG
3001 CCAGGCATCA AATTAAGCAG AAGGCCATCC TGACGGATGG CCTTTTTGCG TTTCTACAAA
3061 CTCTTTCGGT CCGTTGTTTA TTTTTCTAAA TACATTCAAA TATGTATCCG CTCATGAGAC
3121 AATAACCCTG ATAAATGCTT CAATAATATT GAAAAAGGAA GAGTATGAGT ATTCAACATT
3181 TCCGTGTCGC CCTTATTCCC TTTTTTGCGG CATTTTGCCT TCCTGTTTTT GCTCACCCAG
3241 AAACGCTGGT GAAAGTAAAA GATGCTGAAG ATCAGTTGGG TGCACGAGTG GGTTACATCG
3301 AACTGGATCT CAACAGCGGT AAGATCCTTG AGAGTTTTCG CCCCGAAGAA CGTTTCCCAA
3361 TGATGAGCAC TTTTAAAGTT CTGCTATGTG GCGCGGTATT ATCCCGTGTT GACGCCGGGC
3421 AAGAGCAACT CGGTCGCCGC ATACACTATT CTCAGAATGA CTTGGTTGAG TACTCACCAG
3481 TCACAGAAAA GCATCTTACG GATGGCATGA CAGTAAGAGA ATTATGCAGT GCTGCCATAA
3541 CCATGAGTGA TAACACTGCG GCCAACTTAC TTCTGACAAC GATCGGAGGA CCGAAGGAGC
3601 TAACCGCTTT TTTGCACAAC ATGGGGGATC ATGTAACTCG CCTTGATCGT TGGGAACCGG
3661 AGCTGAATGA AGCCATACCA AACGACGAGC GTGACACCAC GATGCCTGTA GCAATGGCAA
3721 CAACGTTGCG CAAACTATTA ACTGGCGAAC TACTTACTCT AGCTTCCCGG CAACAATTAA
3781 TAGACTGGAT GGAGGCGGAT AAAGTTGCAG GACCACTTCT GCGCTCGGCC CTTCCGGCTG
3841 GCTGGTTTAT TGCTGATAAA TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT ATCATTGCAG
3901 CACTGGGGCC AGATGGTAAG CCCTCCCGTA TCGTAGTTAT CTACACGACG GGGAGTCAGG
3961 CAACTATGGA TGAACGAAAT AGACAGATCG CTGAGATAGG TGCCTCACTG ATTAAGCATT
4021 GGTAACTGTC AGACCAAGTT TACTCATATA TACTTTAGAT TGATTTCCTT AGGACTGAGC
4081 GTCAACCCCG TAGAAAAGAT CAAAGGATCT TCTTGAGATC CTTTTTTTCT GCGCGTAATC
4141 TGCTGCTTGC AAACAAAAAA ACCACCGCTA CCAGCGGTGG TTTGTTTGCC GGATCAAGAG
4201 CTACCAACTC TTTTTCCGAA GGTAACTGGC TTCAGCAGAG CGCAGATACC AAATACTGTC
4261 CTTCTAGTGT AGCCGTAGTT AGGCCACCAC TTCAAGAACT CTGTAGCACC GCCTACATAC
4321 CTCGCTCTGC TAATCCTGTT ACCAGTGGCT GCTGCCAGTG GCGATAAGTC GTGTCTTACC
4381 GGGTTGGACT CAAGACGATA GTTACCGGAT AAGGCGCAGC GGTCGGGCTG AACGGGGGGT
4441 TCGTGCACAC AGCCCAGCTT GGAGCGAACG ACCTACACCG AACTGAGATA CCTACAGCGT
4501 GAGCTATGAG AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG CGGACAGGTA TCCGGTAAGC
4561 GGCAGGGTCG GAACAGGAGA GCGCACGAGG GAGCTTCCAG GGGGAAACGC CTGGTATCTT
4621 TATAGTCCTG TCGGGTTTCG CCACCTCTGA CTTGAGCGTC GATTTTTGTG ATGCTCGTCA
4681 GGGGGGCGGA GCCTATGGAA AAACGCCAGC AACGCGGCCT TTTTACGGTT CCTGGCCTTT
4741 TGCTGGCCTT TTGCTCACAT GTTCTTTCCT GCGTTATCCC CTGATTCTGT GGATAACCGT
4801 ATTACCGCCT TTGAGTGAGC TGATACCGCT CGCCGCAGCC GAACGACCGA GCGCAGCGAG
4861 TCAGTGAGCG AGGAAGCGGA AGAGCGCCTG ATGCGGTATT TTCTCCTTAC GCATCTGTGC
4921 GGTATTTCAC ACCGCATATA AGGTGCACTG TGACTGGGTC ATGGCTGCGC CCCGACACCC
4981 GCCAACACCC GCTGACGCGC CCTGACGGGC TTGTCTGCTC CCGGCATCCG CTTACAGACA
5041 AGCTGTGACC GTCTCCGGGA GCTGCATGTG TCAGAGGTTT TCACCGTCAT CACCGAAACG
5101 CGCGAGGCAG CTGCGGTAAA GCTCATCAGC GTGGTCGTGC AGCGATTCAC AGATGTCTGC
5161 CTGTTCATCC GCGTCCAGCT CGTTGAGTTT CTCCAGAAGC GTTAATGTCT GGCTTCTGAT
5221 AAAGCGGGCC ATGTTAAGGG CGGTTTTTTC CTGTTTGGTC ACTGATGCCT CCGTGTAAGG
5281 GGGATTTCTG TTCATGGGGG TAATGATACC GATGAAACGA GAGAGGATGC TCACGATACG
5341 GGTTACTGAT GATGAACATG CCCGGTTACT GGAACGTTGT GAGGGTAAAC AACTGGCGGT
5401 ATGGATGCGG CGGGACCAGA GAAAAATCAC TCAGGGTCAA TGCCAGCGCT TCGTTAATAC
5461 AGATGTAGGT GTTCCACAGG GTAGCCAGCA GCATCCTGCG ATGCAGATCC GGAACATAAT
5521 GGTGCAGGGC GCTGACTTCC GCGTTTCCAG ACTTTACGAA ACACGGAAAC CGAAGACCAT
5581 TCATGTTGTT GCTCAGGTCG CAGACGTTTT GCAGCAGCAG TCGCTTCACG TTCGCTCGCG
5641 TATCGGTGAT TCATTCTGCT AACCAGTAAG GCAACCCCGC CAGCCTAGCC GGGTCCTCAA
5701 CGACAGGAGC ACGATCATGC GCACCCGTGG CCAGGACCCA ACGCTGCCCG AAATT