pLJM1 p18 mito
(Plasmid #26638)

• Depositing Lab: David Sabatini
• Publication: Sancak et al Cell. 2010 Apr 16. 141(2):290-303.

Backbone

• Vector backbone: pLJM1
• Backbone manufacturer: Available at Addgene (#19319)
• Backbone size w/o insert (bp): 7300
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Growth instructions: Grow in XL10Gold or Stbl3 cells to decrease recombination. Growing in SOC also helps.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: p18 mito
• Alt name: p18
• Alt name: c11orf59
• Species: H. sapiens (human)
• Entrez Gene: LAMTOR1 (a.k.a. C11orf59, PDRO, Ragulator1, p18, p27RF-Rho)
• Tag / Fusion Protein:
  • Flag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (unknown if destroyed)
• 3′ cloning site: EcoRI (unknown if destroyed)
• 5′ sequencing primer: CMV-F

Terms and Licenses

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

Depositor Comments

The p18 mito expression
plasmid was generated by cloning of a mutant p18 with amino acids 2–4 changed to alanines into a modified version of the pLKO.1 vector that added, to the C terminus of p18, the mitochondrial localization signal of OMP25
protein.

The order of the construct is the following: Age1-flag-sal-p18(residues 2-4 mutated to alanine, stop condon removed)-GGGGS (2X polylinker)-omp25 seq-stop codon-ecor1