pET23PI-Sce ESARC D218A
(Plasmid #195608)

• Purpose: Expresses PI-SceI (VDE) homing endonuclease/intein from the T7 promoter that contains the D218A active site mutation
• Depositing Lab: Frederick Gimble
• Publication: Gimble et al J Biol Chem. 1995 Mar 17;270(11):5849-56. doi: 10.1074/jbc.270.11.5849.

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: T928C, T929C, A942C
• 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. Contains a D218A mutation in the first LAGLIDADG motif that inactivates the protein.