pET-33b(+) vector (V010984)

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LOCUS       pET-33b(+).        5383 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing proteins that can be 32P-labeled 
            using protein kinase A.
ACCESSION   .
VERSION     .
KEYWORDS    pET-33b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5383)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5383)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5383
                     /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..203
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(206..238)
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(242..256)
                     /codon_start=1
                     /product="site for labeling with protein kinase A"
                     /label=site for labeling with protein kinase A
                     /note="PKA site"
                     /note="The serine is phosphorylated."
                     /translation="RRASV"
     CDS             complement(257..274)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(284..301)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(311..313)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(320..342)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(357..381)
                     /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(382..400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        709..786
                     /label=lacI promoter
     CDS             787..1866
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1882..1903
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2678..2866
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2971..3113
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3299..3887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4009..4821
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4917..5372)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"