pGEX6P1-optimizedNedd4.2F823D
(Plasmid #165105)

• Purpose: bacterial expression of a GST fusion of Human Nedd4.2d F823D
• Depositing Lab: Arthur Haas
• Publication: Todaro et al J Biol Chem. 2018 Nov 23;293(47):18192-18206. doi: 10.1074/jbc.RA118.003716. Epub 2018 Oct 4.

Backbone

• Vector backbone: pGEX6P1
• 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: Nedd4.2d F823D
• Alt name: NEDD4L
• Species: H. sapiens (human)
• Insert Size (bp): 2868
• Mutation: codon optimized for E. coli expression; F823D; mutation for destabilization of trimer formation
• Entrez Gene: NEDD4L (a.k.a. NEDD4-2, NEDD4.2, PVNH7, RSP5, hNEDD4-2)
• Tag / Fusion Protein:
  • GST from plasmid. (N terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning

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.

5' Cloning Site: BamHI (not destroyed), 3' Cloning Site: EcoRI (not destroyed).