mCherry-p62-TIR-AAA
(Plasmid #187901)

• Purpose: Mammalian cell expression of a triple mutant of p62: P228A, E230A, P232A
• Depositing Lab: Erika Holzbaur
• Publication: Harding et al Mol Cell. 2023 Sep 7;83(17):3188-3204.e7. doi: 10.1016/j.molcel.2023.08.005.

Backbone

• Vector backbone: mCherry-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4000
• Total vector size (bp): 6197
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SQSTM1
• Alt name: p62
• Species: H. sapiens (human)
• Insert Size (bp): 2200
• Mutation: Triple alanine mutant: P228A, E230A, P232A
• GenBank ID: NM_003900.5
• Entrez Gene: SQSTM1 (a.k.a. A170, DMRV, EBIAP, FTDALS3, NADGP, OSIL, PDB3, ZIP3, p60, p62, p62B)
• Promoter: CMV
• Tag / Fusion Protein:
  • mCherry (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BstGI (not destroyed)
• 3′ cloning site: XmaI (not destroyed)
• 5′ sequencing primer: pd2.5-mCherry 5'-CACCTACAAGGCCAAGAAGC
• 3′ sequencing primer: pEGFP-C Rev2 5'-CTTTATTTGTAACCATTATAAGCTGC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: S. Martens (University of Vienna); Wurzer et al., eLife 2015

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2022.06.21.496850v1 for bioRxiv preprint.