pcDNA3-mito-APEX
(Plasmid #42607)

• Purpose: APEX targeted to the mitochondrial matrix of mammalian cells. Used for proteomic mapping of mitochondria. This construct has been replaced by APEX2 (construct 49386).
• Depositing Lab: Alice Ting
• Publication: Rhee et al Science. 2013 Jan 31.

Backbone

• Vector backbone: pcDNA3
• Total vector size (bp): 6258
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: mito soybean APEX
• Species: soybean
• Insert Size (bp): 870
• Mutation: K14D, W41F, E112K
• Promoter: CMV
• Tag / Fusion Protein:
  • V5 tag (after mito sequence) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: CMVf
• 3′ sequencing primer: BHGrev

Resource Information

• Supplemental Documents:
  • APEX2 Recommendations from Ting Lab
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This plasmid is for expression of soybean APEX W41F in the mitochondrial matrix under the CMV promoter. In this construct, APEX has a mitochondrial localization peptide (MLATRVFSLVGKRAISTSVCVRAH) fused to its N-terminus.

This plasmid is considered an improved replacement of Addgene plasmid #40309

Also see the following reference:Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy. Martell JD, Deerinck TJ, Sancak Y, Poulos TL, Mootha VK, Sosinsky GE, Ellisman MH, Ting AY. Nat Biotechnol. 2012 Oct 21.(PMID:
23086203)