pMHE2 vector (V004740)

Basic Vector Information

Vector Name:
pMHE2
Antibiotic Resistance:
Streptomycin
Length:
5878 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.

pMHE2 vector Vector Map

pMHE25878 bp60012001800240030003600420048005400T7 promoterlac operatorRBSfusion protein start codon6xHisFactor Xa siteT7 terminatorpBBR1 ReppBBR1 oriVRP4 mob; oriTT4 terminatorSmRT4 terminator

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 ℃.

pMHE2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_30815        5878 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5878)
  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 5878)
  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 5878)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5878)
  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..5878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    226..250
                     /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             265..287
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    294..296
                     /label=fusion protein start codon
                     /note="fusion protein start codon"
     CDS             300..317
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             318..329
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     terminator      442..489
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1123..1782)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1783..2552)
                     /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    2776..3769
                     /note="RP4 mob; oriT"
     regulatory      complement(3859..3974)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4237..5025)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      5761..5876
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"

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