raft-APEX2
(Plasmid #205089)

• Purpose: Expresses lipid-raft targeted APEX2 for proximity proteomics. Vector: pcDNA™4/TO. Insert: lipid raft target sequence (MGCVCSS) + mCherry +V5 (GKPIPNPLLGLDST) +APEX2
• Depositing Lab: Pieta Mattila
• Publication: B cell receptor-induced protein dynamics and the emerging role of SUMOylation revealed by proximity proteomics (unpublished)

Backbone

• Vector backbone: pcDNA™4/TO
• Backbone manufacturer: Thermo Fisher #V102020
• Backbone size w/o insert (bp): 5078
• Total vector size (bp): 6698
• Vector type: Mammalian Expression
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: APEX2 (raft-mCherry-V5 tagged)
• Species: soybean
• Insert Size (bp): 1620
• Tag / Fusion Protein:
  • lipid raft targeting sequence (MGCVCSS), mCherry, V5 (GKPIPNPLLGLDST) (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV
• 3′ sequencing primer: SV40

Resource Information

• A portion of this plasmid was derived from a plasmid made by: APEX cloned originally from pcDNA3-mito-APEX, Addgene Plasmid #42607. Mutated into APEX2 as in: Hung, V., Udeshi, N., Lam, S. et al. Spatially resolved proteomic mapping in living cells with the engineered peroxidase APEX2. Nat Protoc 11, 456–475 (2016). https://doi.org/10.1038/nprot.2016.018

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry