pGAN230
(Plasmid #131122)

• Purpose: (Empty Backbone) reverse yTRAP, RNQ1-aTF reporting on tetR-mKate2
• Depositing Lab: Ahmad Khalil
• Publication: Newby et al Cell. 2017 Nov 2;171(4):966-979.e18. doi: 10.1016/j.cell.2017.09.041. Epub 2017 Oct 19.

Backbone

• Vector backbone: pGAN147
• Backbone manufacturer: Gregory Newby
• Backbone size (bp): 11381
• Modifications to backbone: mNeonGreen replaced with tetR-mKate2
• Vector type: Yeast Expression
• Promoter: SUP35
• Selectable markers: Nourseothricin

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Copy number: High Copy

Cloning Information

• Cloning method: Gateway Cloning

Resource Information

• Supplemental Documents:
  • GAN230-TetR-mKate Prion Sensor plasmid.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This is the empty reverse yTRAP sensor plasmid. To sense aggregation of your favorite protein, clone the coding sequence into a pENTR clone, then perform LR reaction with pGAN230. The resulting expression clone should be linearized with NotI-HF enzyme prior to integration into the yeast genome. The sensor integrates into the HO locus, and can be selected for using nourseothricin. To achieve dual color reporting, combine with pSK221.