pLV UBC APEX2-3xFLAG-IRES-eGFP
(Plasmid #214983)

• Purpose: Cytosolic APEX2 control. Can be used as a C-terminal APEX2 fusion by digestion with NheI and Gibson cloning (see comments).
• Depositing Lab: Wade Harper
• Publication: Hollingsworth et al bioRxiv 2024.04.19.590338

Backbone

• Vector backbone: pLV UBC
• Vector type: Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: APEX2
• Species: Glycine max (soybean)
• Entrez Gene: APX2 (a.k.a. GLYMA_12G073100v4, apx1)
• Promoter: UBC
• Tag / Fusion Protein:
  • 3xFLAG (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TTATATAAGGACGCGCCG

Terms and Licenses

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

Depositor Comments

To clone something N-terminal to APEX2, digest with NheI. Add the following overhangs to forward and reverse primers of the insert: F: 5'-TCCATAGAAGACACCGACGCCACCATG[inset 5' overlap]-3', R: 5'-TCCATAGAAGACACCGACGCCACCATG[Insert 3' overlap reverse complement]-3'. On the 3' side, include a small linker, otherwise there is very little spacer between the insert and APEX2. Additional data, code, and other details related to this work (Hollingsworth et al. 2024) are available at https://harperlab.pubpub.org/pub/nlrp3/.