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pcDNA3 soybean APEX (IMS)
(Plasmid #40309)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 40309 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • 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
  • (Common Sequencing Primers)

Resource Information

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

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pcDNA3 soybean APEX (IMS) was a gift from Alice Ting (Addgene plasmid # 40309)
  • For your References section:

    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. doi: 10.1038/nbt.2375. 10.1038/nbt.2375 PubMed 23086203