pET-51b(+) Ek/LIC vector (V010953)

LOCUS       pET-51b(+)_Ek_LI        5218 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector with an enterokinase site for ligation-independent 
            cloning (LIC) to express proteins tagged with Strep-Tag II.
ACCESSION   .
VERSION     .
KEYWORDS    pET-51b(+) Ek_LIC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5218)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5218)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For LIC, linearize with BseRI and KpnI and treat with T4 DNA 
            polymerase plus dTTP.
FEATURES             Location/Qualifiers
     source          1..5218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(113..139)
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             complement(209..223)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(230..253)
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             complement(260..262)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(269..291)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(306..330)
                     /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)."
     promoter        complement(331..349)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        662..739
                     /label=lacI promoter
     CDS             740..1819
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1835..1856
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2394..2582
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2687..2829
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3015..3603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3684..3776
                     /label=AmpR promoter
     CDS             3777..4634
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(4752..5207)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"