CRY2-INPP5E(D556A)
(Plasmid #79564)

• Purpose: expression of CRY2PHR fused to catalytically dead INPP5E
• Depositing Labs: Pietro De Camilli, Olof Idevall-Hagren
• Publication: Idevall-Hagren et al Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):E2316-23. doi: 10.1073/pnas.1211305109. Epub 2012 Jul 30.

Backbone

• Vector backbone: mCherry-CRY2PHR
• Backbone manufacturer: C. Tucker, University of Colorado Denver
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: INPP5E
• Species: H. sapiens (human)
• Mutation: C-terminal region, aa 214_644, C641A mutation to delete the C-terminal CAAX box and phosphatase dead D556A mutation
• Entrez Gene: INPP5E (a.k.a. CORS1, CPD4, JBTS1, MORMS, PPI5PIV, pharbin)
• Promoter: CMV
• Tag / Fusion Protein:
  • CRY2 (photolyase homology region domain of Arabidopsis thaliana cryptochrome 2) (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PvuI (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: SV40pA-R

Terms and Licenses

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

Depositor Comments

This construct, lacking the N-terminal fluorescent protein, was generated by Nhe1/Not1 digestion to remove mCherry followed by Mungbean nuclease treatment and blunt end ligation.