Str-KDEL_TNF-SBP-RpHLuorin2
(Plasmid #171722)

• Purpose: Synchronize trafficking of TNFa-RpHLuorin2
• Depositing Lab: Geert van den Bogaart
• Publication: Linders et al ACS Chem Biol. 2022 Jan 21;17(1):240-251. doi: 10.1021/acschembio.1c00907. Epub 2022 Jan 10.

Backbone

• Vector backbone: pIRESneo3
• Backbone manufacturer: Clontech
• Modifications to backbone: RUSH TNFα-RpHLuorin2 was generated by replacing EGFP from Str-KDEL_TNF-SBP-EGFP (Addgene plasmid #65278) with synthetic RpHLuorin2 (Genscript) with restriction sites SbfI and BsrGI.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: TNF
• Alt name: DIF, TNF-alpha, TNFA, TNFSF2, TNLG1F
• Species: H. sapiens (human)
• GenBank ID: NM_000594.3
• Entrez Gene: TNF (a.k.a. DIF, TNF-alpha, TNFA, TNFSF2, TNLG1F)
• Promoter: CMV
• Tags / Fusion Proteins:
  • ratiometric pHluorin2 (C terminal on insert)
  • Streptavidin Binding Protein (SBP) (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SbfI (unknown if destroyed)
• 3′ cloning site: BsrGI (unknown if destroyed)
• 5′ sequencing primer: IRES-F
• 3′ sequencing primer: EGFP-N

Resource Information

• A portion of this plasmid was derived from a plasmid made by: TNFα-RpHLuorin2 was generated by replacing EGFP from Str-KDEL_TNF-SBP-EGFP (Addgene plasmid #65278) with synthetic RpHLuorin2 (Genscript)

Terms and Licenses

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

Depositor Comments

Ratiometric pHLuorin2 sequence synthesized by Genscript, original sequence from https://www.scirp.org/journal/paperinformation.aspx?paperid=5043