pYLL201
(Plasmid #53454)

• Purpose: Expresses HA-tagged mouse DNA topoisomerase IIbeta (full length cDNA, in pCS2+ backbone)
• Depositing Labs: Leroy Liu, Lisa Lyu
• Publication: Ban et al Mol Cell Biol. 2013 Oct;33(20):4008-16. doi: 10.1128/MCB.00403-13. Epub 2013 Aug 12.

Backbone

• Vector backbone: pCS2+
• Backbone size w/o insert (bp): 4095
• Total vector size (bp): 9045
• Modifications to backbone: DNA sequence deleted between Cla1 and Xba1 in Polylinker 1.
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: HA-tagged Mus musculus topoisomerase (DNA) II beta (Top2b)
• Alt name: HA-mTop2b
• Alt name: Top2b
• Alt name: topoisomerase (DNA) II beta
• Species: M. musculus (mouse)
• Insert Size (bp): 4873
• GenBank ID: NM_009409
• Entrez Gene: Top2b (a.k.a. D230016L12Rik, Top-2)
• Promoter: CMV
• Tag / Fusion Protein:
  • HA (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Cla1 (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: SP6 primer
• 3′ sequencing primer: T7 primer

Terms and Licenses

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

Depositor Comments

The full length mTop2b cDNA was amplified by RT-PCR using the total RNA isolated from the whole brain of E16.5 mouse embryo. The 5' forward primer contains the Cla1 restriction site and the HA tag, and the 3' reverse primer contains the Xba1 restriction site.

Addgene QC sequencing found an A253V mutation, it should not effect the function of the plasmid.