pMHE5Tc vector (V004736)

Basic Vector Information

Vector Name:
pMHE5Tc
Antibiotic Resistance:
Tetracycline
Length:
6032 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.

pMHE5Tc vector Map

pMHE5Tc6032 bp30060090012001500180021002400270030003300360039004200450048005100540057006000T7 promoterlac operatorRBSfusion protein start codon6xHisFactor Xa siteFLAGStrep-Tag IIT7 terminatorpBBR1 ReppBBR1 oriVRP4 mob; oriTT4 terminatorTcRT4 terminator

pMHE5Tc vector Sequence

LOCUS       40924_30835        6032 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE5Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  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 6032)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6032)
  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..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    206..230
                     /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             245..267
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    274..276
                     /label=fusion protein start codon
                     /note="fusion protein start codon"
     CDS             280..297
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             298..309
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             364..387
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             391..414
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      482..529
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1163..1822)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1823..2592)
                     /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    2816..3809
                     /note="RP4 mob; oriT"
     regulatory      complement(3899..4014)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4323..5510)
                     /label=TcR
                     /note="tetracycline efflux protein"
     regulatory      5895..6010
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"

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