pCoGWA vector (V008323)

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LOCUS       40924_12600        7033 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCoGWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  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  JOURNAL   Unpublished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 7033)
  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 7033)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7033)
  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            "Unpublished"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..7033
                     /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)"
     CDS             106..123
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    139..263
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        288..318
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             372..1028
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1373..1675
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1719..1843)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1921..1939
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1921..1938
                     /regulatory_class="promoter"
     regulatory      1940..1964
                     /regulatory_class="attenuator"
     protein_bind    1940..1964
                     /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      1992..1998
                     /regulatory_class="ribosome_binding_site"
     CDS             2034..2057
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      2098..2145
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2182..2637
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2755..3612)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3613..3705)
                     /label=AmpR promoter
     rep_origin      3786..4374
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4560..4702)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4807..4995)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5533..5554)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5570..6649)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6650..6727)
                     /label=lacI promoter