pFP5370 ss-FIREmate-HA- KDEL-IVS-IRES- TNF-SBP- mCherry-FIREtag
(Plasmid #216696)

• Purpose: Synchronize trafficking of TNFalpha from the ER (RUCH system)
• Depositing Lab: Franck Perez
• Publication: Bottone et al Nat Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8.

Backbone

• Vector backbone: pIRES_neo3
• Backbone manufacturer: Clontech
• Total vector size (bp): 6587
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: FIREmate-KDEL
• Insert Size (bp): 528
• Promoter: CMV

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: T7_forward

Gene/Insert 2

• Gene/Insert name: Tumor Necrosis Factor alpha (TNFa)
• Species: H. sapiens (human)
• Insert Size (bp): 1761
• Promoter: CMV
• Tags / Fusion Proteins:
  • Streptavidin Binding Protein (SBP) (C terminal on insert)
  • mCherry (C terminal on insert)
  • FIREtag (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: AscI (not destroyed)
• 3′ cloning site: PacI (not destroyed)
• 5′ sequencing primer: IRES-F

Resource Information

• Supplemental Documents:
  • 216696.gbk

Terms and Licenses

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