pCo0GWC vector (V008325)

Basic Vector Information

Vector Name:
pCo0GWC
Antibiotic Resistance:
Chloramphenicol
Length:
5600 bp
Type:
Expression vector
Replication origin:
p15A ori
Source/Author:
Busso D.

pCo0GWC vector Map

pCo0GWC5600 bp60012001800240030003600420048005400T7 promoterlac operatorRBSattR1lac UV5 promoterCmRccdBattR2T7 promoterregulatoryregulatoryFLAGT7 terminatorCmRcat promoterp15A oriCAP binding sitelacIlacI promoter

pCo0GWC vector Sequence

LOCUS       40924_12585        5600 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCo0GWC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5600)
  AUTHORS   Busso D.
  TITLE     Construction of a set of Gateway-based co-expression vectors for 
            rapid cloning and multi-protein production in Escherichia coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5600)
  AUTHORS   Busso D.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2012) Molecular Biology, IGBMC, 1 rue Laurent 
            Fries, Illkirch, Alsace 67404, France
REFERENCE   3  (bases 1 to 5600)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5600)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUN-2012) Molecular Biology, IGBMC, 1 rue Laurent Fries, 
            Illkirch, Alsace 67404, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    26..50
                     /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             65..87
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    118..242
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        267..297
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             351..1007
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1352..1654
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1698..1822)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1900..1918
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1900..1917
                     /regulatory_class="promoter"
     regulatory      1919..1943
                     /regulatory_class="attenuator"
     protein_bind    1919..1943
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     regulatory      1971..1977
                     /regulatory_class="ribosome_binding_site"
     CDS             2013..2036
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      2077..2124
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2347..3003)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3004..3106)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(3632..4176)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(4367..4388)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4404..5483)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5484..5561)
                     /label=lacI promoter

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