| 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: | -- |
买家导航
pMXB10载体质粒基本信息
pMXB10质粒图谱载体图谱和pMXB10载体序列质粒序列多克隆位点信息
pMXB10质粒载体简介
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.
pMXB10质粒序列载体序列
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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pMXB10质粒载体应用举例
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