pGEX-KG-D4H*-mCherry
(Plasmid #134604)

• Purpose: fluorescent cholesterol-binding probe
• Depositing Lab: Roberto Zoncu
• Publication: Lim et al Nat Cell Biol. 2019 Oct;21(10):1206-1218. doi: 10.1038/s41556-019-0391-5. Epub 2019 Sep 23.

Backbone

• Vector backbone: pGEX-KG
• Backbone size w/o insert (bp): 5006
• Total vector size (bp): 6203
• Vector type: Bacterial 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

• Gene/Insert name: D4H*
• Species: Synthetic; Clostridium perfringens
• Insert Size (bp): 333
• Mutation: Y415A and A463W
• GenBank ID: NC_008261
• Tags / Fusion Proteins:
  • GST (N terminal on backbone)
  • mCherry (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: HpaI (destroyed during cloning)
• 5′ sequencing primer: GTATTGAAGCTATCCCAC
• 3′ sequencing primer: AGACAAGCTGTGACCGTC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: D4H–mCherry cDNA was obtained from Gregory D. Fairn (University of Toronto) and inserted into the pGEX-KG vector. The construct was renamed to D4H*–mCherry as two mutations, Y415A and A463W, were introduced to improve the sensitivity of cholesterol binding, in addition to the D434S mutation in the original D4H cDNA.
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

https://benchling.com/s/seq-qYjuRHB1UTwPC8BVFbvK