pGEX4T-3-6AOptParkin
(Plasmid #165107)

• Purpose: bacterial expression of a GST fusion of e.coli optimized Parkin with 6 alanine linker between the GST and the inserted Parkin gene
• Depositing Lab: Arthur Haas
• Publication: Haas Lab plasmids (unpublished)

Backbone

• Vector backbone: pGEX4T-3
• Backbone manufacturer: GE Healthcare
• Backbone size w/o insert (bp): 4900
• 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: Parkin isoform 1
• Alt name: PARK2
• Species: H. sapiens (human)
• Insert Size (bp): 1413
• Mutation: codon optimized for E. coli expression
• Entrez Gene: PRKN (a.k.a. AR-JP, LPRS2, PARK2, PDJ)
• Tag / Fusion Protein:
  • GST-6-alanine (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Sal1 (not destroyed)
• 3′ cloning site: Not1 (not destroyed)

Terms and Licenses

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

Depositor Comments

Best yields for protein purification were found in BL21 (DE) cells using an overnight induction at 16 degrees C.