pGEX-E6AP Hect
(Plasmid #165102)

• Purpose: bacterial expression of a GST fusion of Human E6AP Hect domain
• Depositing Lab: Arthur Haas
• Publication: Ronchi et al Methods Mol Biol. 2012;832:197-218. doi: 10.1007/978-1-61779-474-2_14.

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: E6AP Hect
• Alt name: UBE3A Hect
• Species: H. sapiens (human)
• Insert Size (bp): 731
• Mutation: E6AP Hect domain Amino acids 495-738 (5'QLNPY-LICG3')
• 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.