pET-50b(+) vector (V010955)

LOCUS       pET-50b(+).        6733 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing proteins with a cleavable N-terminal
            NusA tag and a cleavable C-terminal S-tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-50b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6733)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6733
                     /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(168..212)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(213..230)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     misc_feature    234..308
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(309..332)
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             complement(342..359)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(375..1859)
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
     CDS             complement(1866..1883)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(1893..1895)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(1903..1925)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1940..1964)
                     /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(1965..1983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2296..2373
                     /label=lacI promoter
     CDS             2374..3453
                     /label=lacI
                     /note="lac repressor"
     protein_bind    3469..3490
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4028..4216
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4321..4463
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4649..5237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             5359..6171
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(6267..6722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"