TRIPZ-CTID195-GSQ-UBnc-PURO
(Plasmid #208037)

• Purpose: Enables inducible expression of C-terminal Split-TurboID-fused UBnc, to perform Ubiquitin-ID; nc = non-cleavable mutation; selection with puromycin
• Depositing Labs: Rosa Barrio, James Sutherland
• Publication: Barroso-Gomila et al Nat Commun. 2021 Nov 18;12(1):6671. doi: 10.1038/s41467-021-26807-6.

Backbone

• Vector backbone: TRIPZ
• Backbone manufacturer: Open Biosystems
• Vector type: Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ubnc
• Species: H. sapiens (human)
• Mutation: L73P Mutation near C-terminus suppresses cleavage by SENPs/DUBs
• Entrez Gene: UBC (a.k.a. HMG20)
• Promoter: tetO/UBC
• Tag / Fusion Protein:
  • Myc, C-terminal Split-TurboID (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• A portion of this plasmid was derived from a plasmid made by: TurboID fragments derived from Addgene ID # 107169 (Ting Lab).

Terms and Licenses

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

Depositor Comments

Lentiviral packaging of TRIPZ is more efficient with TAT expression (e.g using pcDNA1-TatHIV Addgene ID: 138478) and 2nd generation gag/pol/env combination (e.g. using PSPAX2 Addgene ID: 12260 and pMD2.G Addgene ID: 12259); UBnc is excisable using EcoR1-Mlu1.