| 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: | -- |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
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
//
