pRluc-LC3G120A
(Plasmid #105003)

• Purpose: Mammalian expression of Rluc-LC3G120A. Can be used for measuring autophagic flux.
• Depositing Lab: Marja Jããttelã
• Publication: Farkas et al Methods Enzymol. 2017;588:1-13. doi: 10.1016/bs.mie.2016.09.072. Epub 2016 Nov 12.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4000
• Total vector size (bp): 5300
• Modifications to backbone: Rluc-LC3G120A ORF inserted instead of the EGFP ORF.
• Vector type: Mammalian Expression, Luciferase
• 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: Unknown

Gene/Insert

• Gene/Insert name: LC3B
• Alt name: MAP1LC3B
• Species: R. norvegicus (rat)
• Mutation: Glycine 120 mutated to alanine, prevents lipidation of the protein and thereby its binding to autophagosomal membranes
• Entrez Gene: Map1lc3b (a.k.a. LC3B, Map1lc3, Mpl3, zbs559)
• Tag / Fusion Protein:
  • Rluc with C124A substitution (N terminal on insert)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The gene was originally cloned in Tamotsu Yoshimori Laboratory, reference Kabeya et al. 2000 and addgene ID 21073.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Renilla Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry