pET-30b(+) vector (V010994)

LOCUS       pET-30b(+).        5421 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with an enterokinase site. For other reading frames, use pET-30a(+) 
            or pET-30c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-30b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5421)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5421)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5421
                     /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(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    158..216
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(218..232)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(248..292)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(299..316)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(326..343)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(344..346)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(354..376)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(391..415)
                     /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(416..434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        747..824
                     /label=lacI promoter
     CDS             825..1904
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1920..1941
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2716..2904
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3009..3151
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3337..3925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4047..4859
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4955..5410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"