Basic Vector Information
- Vector Name:
- pMHE5
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5938 bp
- Type:
- Broad host range expression vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Fodor BD, Kovacs AT, Csaki R, Hunyadi-Gulyas E, Klement E, Maroti G, Meszaros LS, Medzihradszky KF, Rakhely G, Kovacs KL.
pMHE5 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pMHE5 vector Sequence
LOCUS 40924_30830 5938 bp DNA circular SYN 18-DEC-2018
DEFINITION Broad host range expression vector pMHE5, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5938)
AUTHORS Fodor BD, Kovacs AT, Csaki R, Hunyadi-Gulyas E, Klement E, Maroti G,
Meszaros LS, Medzihradszky KF, Rakhely G, Kovacs KL.
TITLE Modular broad-host-range expression vectors for single-protein and
protein complex purification
JOURNAL Appl. Environ. Microbiol. 70 (2), 712-721 (2004)
PUBMED 14766546
REFERENCE 2 (bases 1 to 5938)
AUTHORS Fodor BD.
TITLE Direct Submission
JOURNAL Submitted (16-MAY-2003) Biophysics, BRC Hungarian Academy of
Sciences, Temesvari krt., Szeged, Csongrad 6701, Hungary
REFERENCE 3 (bases 1 to 5938)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5938)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2004"; volume: "70"; issue: "2"; pages:
"712-721"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences,
Temesvari krt., Szeged, Csongrad 6701, Hungary"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5938
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 193..211
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 212..236
/label=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)."
RBS 251..273
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 280..282
/label=fusion protein start codon
/note="fusion protein start codon"
CDS 286..303
/label=6xHis
/note="6xHis affinity tag"
CDS 304..315
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
CDS 370..393
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 397..420
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
terminator 488..535
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1169..1828)
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
rep_origin complement(1829..2598)
/direction=LEFT
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
misc_feature 2822..3815
/note="RP4 mob; oriT"
regulatory complement(3905..4020)
/label=T4 terminator
/note="T4 terminator"
/regulatory_class="terminator"
CDS complement(4283..5071)
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
regulatory 5807..5922
/label=T4 terminator
/note="T4 terminator"
/regulatory_class="terminator"
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