pET-41 Ek/LIC vector (V010976)

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LOCUS       pET-41_Ek_LIC.        5947 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for ligation-independent cloning (LIC) to express 
            GST-tagged proteins with an enterokinase site.
ACCESSION   .
VERSION     .
KEYWORDS    pET-41 Ek_LIC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5947)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For LIC, linearize with BseRI and treat with T4 DNA polymerase plus 
            dTTP.
FEATURES             Location/Qualifiers
     source          1..5947
                     /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(150..173)
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    174..259
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(279..293)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(324..368)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(384..401)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(411..428)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(456..1109)
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     RBS             complement(1117..1139)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1154..1178)
                     /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(1179..1197)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1510..1587
                     /label=lacI promoter
     CDS             1588..2667
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2683..2704
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3242..3430
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3535..3677
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3863..4451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4573..5385
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(5481..5936)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"