pMal- p5X Plasmid

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  • Model: PVT0411
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pMal-p5X

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pMal-p5X,Plasmid pMal-p5X,pMal-p5X vector

 

pMal-p5X Information

Promoter: Tac

Replicon: ColE1 ori

Terminator: rrnB T1 Terminator

Plasmid classification: Escherichia coli vector; pMal series expression plasmid

Plasmid size: 5752bp

Plasmid label: N-MBP

Prokaryotic resistance: ampicillin Amp

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic, LB

Expression host: BL21 (DE3)

Culture conditions: 37 centigrade, aerobic, LB

Inducement: IPTG or lactose and its analogues

5'sequencing primers: Based on sequence design

3'sequencing primers: Based on sequence design

Remark: fusion expression of maltose binding protein MBP, protein located in the cell cycle

 

 

pMal-p5X Description

pMal-p5X-plasmid

 

 

pMal-p5X Sequence

LOCUS       Exported                5752 bp ds-DNA     circular SYN 19-09-2014
DEFINITION  Bacterial vector for inducible periplasmic expression of
            maltose-binding protein (MBP) fusions with a Factor Xa cleavage
            site.
ACCESSION   .
VERSION     .
KEYWORDS    pMAL-p5X
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  AUTHORS   Riggs P.
  TITLE     Expression and purification of maltose-binding protein fusions.
  JOURNAL   Curr Protoc Mol Biol 2001;Chapter 16:Unit16.6.
  PUBMED    18265133
REFERENCE   2  (bases 1 to 5752)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
  JOURNAL   Exported 2016-09-21 from SnapGene Viewer 2.8.1
             
FEATURES             Location/Qualifiers
     source          1..5752
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        3..80
                     /gene="lacI (mutant)"
                     /note="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             81..1163
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    1406..1434
                     /note="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1528..2703
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /note="MBP"
                     /note="This gene encodes a signal sequence that can direct
                     MBP to the bacterial periplasm."
                     /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             2752..2763
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     misc_feature    2759..2837
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      2838..2924
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3016..3043
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3071..3161
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             3162..4022
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
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