pSK955 pcDNA5-FRT-TO-hTOP2A-2xFLAG
(Plasmid #260231)

• Purpose: human TOP2A WT with C-terminal 2xFLAG DOX-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: pcDNA5/FRT/TO
• Backbone manufacturer: Invitrogen/Thermo Fisher Scientific
• Backbone size w/o insert (bp): 5130
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

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: Low Copy

Gene/Insert

• Gene/Insert name: TOP2A
• Alt name: hTOP2A; DNA topoisomerase II alpha
• Species: H. sapiens (human)
• Insert Size (bp): 4650
• Entrez Gene: TOP2A (a.k.a. TOP2, TOP2alpha, TOPIIA, TP2A)
• Promoter: CMV/TetO promoter
• Tag / Fusion Protein:
  • 2xFLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: ApaI (not destroyed)

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

Flp-In/T-REx pcDNA5/FRT/TO construct for inducible hTOP2A-2xFLAG expression. 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. In-Fusion cloning; vector linearized by restriction digestion. Please visit https://doi.org/10.64898/2026.05.09.723998 for bioRxiv preprint.