pSK815 pRS415 pCUP1-yTOP2-F1025Y,R1128G-3xFLAG
(Plasmid #260230)

• Purpose: yeast TOP2-F1025Y, R1128G with C-terminal 3xFLAG copper-inducible expression
• Depositing Lab: Scott Keeney
• Publication: Ontoso et al bioRxiv [Preprint]. 2026 May 11:2026.05.09.723998. doi: 10.64898/2026.05.09.723998.

Backbone

• Vector backbone: pRS415
• Backbone size w/o insert (bp): 6219
• Vector type: Yeast Expression
• Selectable markers: LEU2

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: Grow in NEB Stable E. coli at 30 C; minimize bacterial propagation and verify sequence after expansion because TOP2 plasmids, especially mutant constructs, can acquire secondary mutations.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TOP2
• Alt name: yTOP2; DNA topoisomerase II; YNL088W
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 4356
• Mutation: F1025Y; R1128G
• Entrez Gene: TOP2 (a.k.a. YNL088W, TOR3, TRF3)
• Promoter: CUP1 promoter
• Tag / Fusion Protein:
  • 3xFLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SacII (not destroyed)
• 5′ sequencing primer: M13 fwd
• 3′ sequencing primer: M13 rev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Synthetic DNA; sequence verified

Terms and Licenses

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

Depositor Comments

pRS415 CEN/ARS LEU2 yeast expression plasmid with CUP1 promoter-driven yTOP2-F1025Y,R1128G-3xFLAG. Grow in NEB Stable E. coli at 30 C; minimize bacterial propagation and verify sequence after expansion because TOP2 plasmids, especially mutant constructs, can acquire secondary mutations. Please visit https://doi.org/10.64898/2026.05.09.723998 for bioRxiv preprint.