pET104-DEST vector (V011797)

LOCUS       pET104-DEST.        7618 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for high-level inducible expression of
            biotinylated proteins in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    pET104-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7618)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7618)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             297..299
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             300..515
                     /label=BioEase(TM) tag
                     /note="biotinylated sequence derived from the C-terminus of
                     the alpha-subunit of Klebsiella pneumoniae oxaloacetate 
                     decarboxylase"
     CDS             528..551
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
     protein_bind    561..685
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        710..740
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             794..1450
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1774..2076
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2120..2244)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2330..2377
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2701..2805
                     /label=AmpR promoter
     CDS             2806..3663
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3837..4425
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4611..4753)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4858..5046)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(6324..6345)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6361..7440)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7441..7518)
                     /label=lacI promoter