pcDNA3 soybean APEX (IMS)
(Plasmid #40309)

• Depositing Lab: Alice Ting
• Publication: Martell et al Nat Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375.

Backbone

• Vector backbone: pcDNA3
• Total vector size (bp): 6390
• 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: soybean APEX
• Species: soybean
• Insert Size (bp): 747
• Mutation: K14D, W41F, E112K mutations
• Promoter: CMV
• Tags / Fusion Proteins:
  • LACTB IMS localization peptide (N terminal on insert)
  • V5 (internal) (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: T7
• 3′ sequencing primer: BGHrev

Resource Information

• Supplemental Documents:
  • APEX2 Recommendations from Ting Lab

Terms and Licenses

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

Depositor Comments

This gene is for expression of soybean APEX in the mitochondrial intermembrane space (IMS) of mammalian cells under the CMV promoter. The APEX gene has the LACTB IMS localization peptide fused to its N-terminus. Soybean APEX has three mutations (K14D, W41F, E112K) relative to wild-type soybean ascorbate peroxidase. Soybean APEX is 90% sequence identical to pea APEX and performs very similarly in cells, but we have some in vitro evidence that soybean APEX is more monomeric than pea APEX. Note that soybean APEX is less characterized, and pea APEX is the focus of Martell et. al. NBT 2012.

THE TING LAB HAS DISCONTINUED THIS PLASMID AND ENCOURAGES SCIENTISTS TO USE THE FOLLOWING UPDATED VERSION: www.addgene.org/42607