mCherry-CRY2-P14P5K
(Plasmid #79570)

• Purpose: expression of mCherry-tagged CRY2PHR fused to iSH2 domain of mouse Pip5k1c
• Depositing Lab: Olof Idevall-Hagren
• Publication: Xie et al Cell Chem Biol. 2016 Jul 21;23(7):816-26. doi: 10.1016/j.chembiol.2016.06.009.

Backbone

• Vector backbone: pEGFP
• 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: Pip5k1c
• Species: M. musculus (mouse)
• Mutation: R445E and R446E; truncation of amino acids 636-661 (please see depositor comments below)
• Entrez Gene: Pip5k1c (a.k.a. AI115456, AI835305, Pip5kIgamma)
• Promoter: CMV
• Tags / Fusion Proteins:
  • mCherry (N terminal on backbone)
  • 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: mCherry-F
• 3′ sequencing primer: SV40pA-R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Three modifications have been made to PIP5K1g (NM008844): R445E and R446E mutations to favour cytosolic distribution and truncation of amino acids 636-661 to prevent plasma membrane localisation.