pMal-p2G Search name
pMal-p2G,Plasmid pMal-p2G,pMal-p2G vector
pMal-p2G information
Promoter: Tac
Replicon: ColE1 ori
Terminator: rrnB T1 Terminator
Plasmid classification: Escherichia coli vector; pMal series expression plasmid
Plasmid size: 6724bp
Plasmid tagging: N-MBP, N-Genenase I
Prokaryotic resistance: ampicillin Amp
Clonal strain: DH5 alpha
Culture conditions: 37 C, aerobic, LB
Expression host: BL21 (DE3)
Culture conditions: 37 C, aerobic, LB
Induction: IPTG or lactose and its analogues.
5'primers: MalE primers: 5-GGTCGTCAGACTGTCGATGAAGCC-3; MBP-F: 5-gatgaagccctgaaagacgcgcag-3
3'sequencing primers: pBad-5 5-gatttaatctgtatcagg-3; M13-F: 5-TGTAAAACGACGGCCAGT-3
Remarks: fusion expression of maltose binding protein MBP, protein located in periplasmic.
pMal-p2G Description
PMAL system is an efficient protein fusion expression and purification system. The pMAL vector contains E. coli malE gene encoding maltose Binding Protein (MBP), and its downstream polyclonal site facilitates gene insertion and expresses the fusion protein with MBP at the N-terminal. The cloned gene was highly expressed by "tac" strong promoter and malE translation initiation signal, and the fusion protein was further purified by one-step affinity purification with Amylose column using MBP affinity to maltose.
pMal-p2G Multiple cloning site
pMal-p2G Sequence
LOCUS Exported 6724 bp ds-DNA circular SYN 13-08-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMal-p2G
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6724)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-13 from SnapGene Viewer 2.8.1
FEATURES Location/Qualifiers
source 1..6724
/organism="synthetic DNA construct"
/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
KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
CDS 2752..2769
/codon_start=1
/product="recognition and cleavage site for Genenase(TM) I,
a subtilisin BPN' variant engineered for increased
substrate specificity (Carter and Wells, 1987)"
/note="Genenase(TM) I site"
/translation="PGAAHY"
primer_bind complement(2814..2830)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
terminator 3179..3265
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3357..3384
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3404..3495
/gene="bla"
/note="AmpR promoter"
CDS 3496..4356
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4398..4911)
温馨提示:不可用于临床ZL。