Tmem98-mCherry
(Plasmid #124505)

• Purpose: Expresses mCherry-tagged mouse TMEM98 in mammalian cells (C-terminal tag)
• Depositing Lab: Renee van Amerongen
• Publication: van der Wal et al PLoS One. 2020 Jan 21;15(1):e0227435. doi: 10.1371/journal.pone.0227435. eCollection 2020.

Backbone

• Vector backbone: pAC-N2
• Backbone size w/o insert (bp): 4500
• Total vector size (bp): 5415
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Tmem98
• Species: M. musculus (mouse)
• Insert Size (bp): 770
• Mutation: stop codon was removed to allow fusion with sGFP2 at the C-terminus
• Entrez Gene: Tmem98 (a.k.a. 6530411B15Rik, AI463522)
• Promoter: CMV
• Tag / Fusion Protein:
  • mCherry (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: SV40 polyA

Terms and Licenses

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

Depositor Comments

The full length Tmem98 coding sequence was initially fused to AC-GFP (Clontech, pAC-GFP-N1 vector).

To generate an sGFP2 fusion, the pAC-GFP-Tmem98 construct was cut with BamHI and BsrgI (to release AC-GFP), after which a BamHI-BsrgI sGFP2 fragment was re-inserted.

The mCherry tagged variant was generated by exchanging an AgeI/BsrgI sGFP2 fragment for an AgeI/BsrgI mTurquoise fragment. Please visit https://www.biorxiv.org/content/10.1101/512426v1 for BioRxiv preprint