Lenti-EFS-BirAopt-GSQ-RNF4ca-P2A-BLAST
(Plasmid #208047)

• Purpose: Enables constitutive expression of N-terminal BirAopt-fused RNF4ca (catalytically inactive RING mutant); to perform bioE3; selection with blasticidin
• Depositing Labs: Rosa Barrio, James Sutherland
• Publication: Barroso-Gomila et al Nat Commun. 2023 Nov 23;14(1):7656. doi: 10.1038/s41467-023-43326-8.

Backbone

• Vector backbone: Derived from Addgene ID# 52962 (Zhang Lab).
• Vector type: Lentiviral
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: RNF4
• Species: H. sapiens (human)
• Mutation: Cys>Ala mutations in conserved RING domain
• Entrez Gene: RNF4 (a.k.a. RES4-26, SLX5, SNURF)
• Promoter: EFS
• Tag / Fusion Protein:
  • BirAopt (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Human codon-optimized BirA was a gift from Oryzko Lab (PMID 15707685). Lenti-EFS/P2A-blast backbone derived from Addgene ID# 52962 (Zhang Lab).

Terms and Licenses

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

Depositor Comments

Control for RNF4 BioE3 experiments; contains mutations in the conserved RING domain. RNF4ca is excisable using EcoR1-Not1.