pET PI-SceI C-His K301A
(Plasmid #195611)

• Purpose: Expresses PI-SceI (VDE) homing endonuclease/intein from the T7 promoter that contains the K301A active site mutation
• Depositing Lab: Frederick Gimble
• Publication: He et al J Biol Chem. 1998 Feb 20;273(8):4607-15. doi: 10.1074/jbc.273.8.4607.

Backbone

• Vector backbone: pET-23a
• Backbone manufacturer: Novagen
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: PI-SceI
• Alt name: VDE
• Species: S. cerevisiae (budding yeast)
• Mutation: A822G, A823C, A824T
• Promoter: T7

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Expresses the excised intein of the VMA1 gene product including amino-terminal methionine-cysteine residues. Contains silent AvrII and RsrII sites near the borders of the N-terminal LAGLIDADG motif and silent EaeI and SnaBI sites near the borders of the C-terminal motif. Also contains silent mutations that introduce ApaI, BssHII, BstEII, BsiWI, MluI and SpeI sites. Expresses PI-SceI containing a carboxyl-terminal histidine tag and a K301A mutation in the PI-SceI ORF.