pRSF-2 Ek/LIC vector (V010924)

LOCUS       pRSF-2_Ek_LIC.        3609 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector with an RSF 1030 origin for ligation-independent 
            cloning (LIC) to express 6xHis- and S-tagged proteins with an 
            enterokinase site.
ACCESSION   .
VERSION     .
KEYWORDS    pRSF-2 Ek_LIC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3609)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3609)
  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..3609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             71..73
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             77..94
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             98..112
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             146..190
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      242..289
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(522..1334)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1335..1426)
                     /label=AmpR promoter
     rep_origin      complement(1442..2191)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(2353..2374)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2390..3469)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3470..3547)
                     /label=lacI promoter
     promoter        3593..3609
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"