pET-29a(+) vector (V011002)

Basic Vector Information

      • Vector Name:
      • pET-29a(+)
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 5371 bp
      • Type:
      • pET & Duet Vectors (Novagen)
      • Source/Author:
      • Novagen (EMD Millipore)
      • Copy Number:
      • High copy number

pET-29a(+) vector Vector Map

pET-29a(+)5371 bp6001200180024003000360042004800T7 terminator6xHisMCSthrombin siteS-TagATGRBSlac operatorT7 promoterlacI promoterlacICAP binding siteropbomoriKanRf1 ori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pET-29a(+) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pET-29a(+).        5371 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with a thrombin site. For other reading frames, use pET-29b(+) or 
            pET-29c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-29a(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5371)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5371)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5371
                     /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(216..233)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(249..293)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(294..296)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(304..326)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(341..365)
                     /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(366..384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        697..774
                     /label=lacI promoter
     CDS             775..1854
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1870..1891
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2666..2854
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2959..3101
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3287..3875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3997..4809
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4905..5360)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"

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