PERK 7: PERK.K618A.9E10.pBABE
(Plasmid #21820)

• Depositing Lab: David Ron
• Publication: PERK constructs (unpublished)

Backbone

• Vector backbone: pBABE-puro
• Backbone manufacturer: Plasmid available at Addgene (#1764)
• Backbone size w/o insert (bp): 5150
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl2
• Growth instructions: We have found that the PERK cDNA confers instability (low replication ability) to plasmids. The plasmid will be provided in the STBL2 bacterial strain (gibco) which helps alleviate this problem. To recover the plasmid, stick a sterile toothpick into the stab and streak on an LB amp plate and grow at 30C. Pick an individual colony and grow in LB amp at 30C for the highest yield of plasmid.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: PERK K618A
• Alt name: Eif2ak3 eukaryotic translation initiation factor 2 alpha kinase 3
• Species: M. musculus (mouse)
• Insert Size (bp): 3500
• Mutation: kinase dead K618A mutant
• Entrez Gene: Eif2ak3 (a.k.a. Pek, Perk)
• Tag / Fusion Protein:
  • Myc (9E10) (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: pBABE 5'
• 3′ sequencing primer: pBABE 3'

Terms and Licenses

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

Depositor Comments

9E10-tagged mouse PERK K618A mutant in the pro-retroviral vector pBABEpuro

Mammalian expression plasmid encoding kinase dead K618A mutant mouse PERK fused to 9E10 epitope at C-term.

Note: Addgene has yet to be able to sequence the C-terminal Myc tag, therefore we cannot guarantee that is present.