pMHE7Tc vector (V004732)

Basic Vector Information

Vector Name:
pMHE7Tc
Antibiotic Resistance:
Tetracycline
Length:
6014 bp
Type:
Broad host range expression vector
Replication origin:
pBBR1 oriV
Source/Author:
Fodor BD.

pMHE7Tc vector Vector Map

pMHE7Tc6014 bp30060090012001500180021002400270030003300360039004200450048005100540057006000T7 promoterlac operatorRBSstart codon of fusion proteinFLAGStrep-Tag IIFactor Xa siteT7 terminatorpBBR1 ReppBBR1 oriVRP4 mob, oriTT4 terminatorTcRT4 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 ℃.

pMHE7Tc vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_30855        6014 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE7Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  AUTHORS   Fodor BD.
  TITLE     Modular broad-host-range expression vectors for the purification of 
            proteins and protein complexes and their application
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6014)
  AUTHORS   Fodor BD.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAY-2003) Biophysics, BRC Hungarian Academy of 
            Sciences, Temesvari, Szeged, Csongrad 6701, Hungary
REFERENCE   3  (bases 1 to 6014)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6014)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences, 
            Temesvari, Szeged, Csongrad 6701, Hungary"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        189..207
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    208..232
                     /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             247..269
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    276..278
                     /label=start codon of fusion protein
                     /note="start codon of fusion protein"
     CDS             282..305
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             318..341
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             342..353
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     terminator      466..513
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1147..1806)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1807..2576)
                     /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    2800..3793
                     /label=RP4 mob, oriT
                     /note="RP4 mob, oriT"
     regulatory      complement(3883..3998)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4307..5494)
                     /label=TcR
                     /note="tetracycline efflux protein"
     regulatory      5879..5994
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"

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