PERK 4: PERKKD-pGEX4T-1
(Plasmid #21817)

• Depositing Lab: David Ron
• Publication: Harding et al Nature. 1999 Jan 21. 397(6716):271-4.

Backbone

• Vector backbone: pGEX-4T-1
• Backbone manufacturer: Amersham
• Backbone size w/o insert (bp): 5000
• Vector type: Bacterial Expression

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 (537-1114)
• Alt name: Eif2ak3 eukaryotic translation initiation factor 2 alpha kinase 3
• Species: M. musculus (mouse)
• Insert Size (bp): 1700
• Mutation: PERK Kinase Domain. Amino acids 537-1114.
• Entrez Gene: Eif2ak3 (a.k.a. Pek, Perk)
• Tags / Fusion Proteins:
  • GST (N terminal on backbone)
  • Myc (9E10) (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: pGEX-F

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

Mouse PERK (537-1114), wild-type, 9E10-tagged at C-terminus, fused to GST, bacterial expression vector, pGEX4T-1

Sequencing found a S1109T mutation. There is no evidence that the mutation affects activity.