pCOLA-3-DEST vector (V011842)

LOCUS       pCOLA-3-DEST.        5134 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) Nova bacterial destination vector with a ColA origin for 
            expressing proteins tagged at the N-terminus with Strep-Tag II.
ACCESSION   .
VERSION     .
KEYWORDS    pCOLA-3-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5134)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5134)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(227..1039)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1040..1131)
                     /label=AmpR promoter
     rep_origin      complement(1149..1784)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     protein_bind    complement(1948..1969)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1985..3064)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3065..3142)
                     /label=lacI promoter
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3207..3231
                     /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             3246..3268
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3275..3277
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             3284..3307
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     protein_bind    3317..3441
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3466..3496
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3550..4206
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4551..4853
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4897..5021)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5023..5040
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      5081..5128
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"