pGEX4T-1-Optimized E6AP
(Plasmid #165100)

• Purpose: ecoli optimized expression of GST-fusion of human E6AP
• Depositing Lab: Arthur Haas
• Publication: Ronchi et al J Biol Chem. 2013 Apr 12;288(15):10349-60. doi: 10.1074/jbc.M113.458059. Epub 2013 Feb 25.

Backbone

• Vector backbone: pGEX4T1
• 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: E6AP isoform 3
• Alt name: UBE3A isoform 3
• Species: H. sapiens (human)
• Insert Size (bp): 2620
• Mutation: codon optimized for E. coli expression
• Entrez Gene: UBE3A (a.k.a. ANCR, AS, E6-AP, EPVE6AP, HPVE6A, PIX1)
• Tag / Fusion Protein:
  • GST from plasmid. (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR1 (not destroyed)
• 3′ cloning site: Xho1 (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.