pB6-Gfap-T2A-tdTomato-FNF-DTA
(Plasmid #140271)

• Purpose: B6J Gfap-T2A-tdTomato allele targeting vector, low copy number
• Depositing Lab: Mario Capecchi
• Publication: Miscellaneous gene targeting vector plasmids (unpublished)

Backbone

• Vector backbone: pDTA-5
• Backbone manufacturer: Hyung-song Nam, Capecchi laboratory
• Vector type: Mammalian Expression, Mouse Targeting
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH10B
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Gfap
• Species: M. musculus (mouse)
• Entrez Gene: Gfap
• Tag / Fusion Protein:
  • C-terminal fusion of T2A-tdTomato (C terminal on insert)

Resource Information

• Supplemental Documents:
  • pGfap-T2A-tdTomato-FNF-DTA5.gb
• A portion of this plasmid was derived from a plasmid made by: tdTomato is from Roger Tsien. FRT-Neo-FRT is from pFNF (#22687).

Terms and Licenses

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

Depositor Comments

A targeting vector to knock-in T2A-tdTomato into mouse Gfap protein isoform 2 (also known as Gfap-alpha). This is the most abundant isoform in the mouse brain. Of note, the Gfap protein transcript variant 2 transcript level is high in the SVZ, an adult neural stem cell niche. See Kamphuis et al., 2012 (https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0042823)

Homology arms subcloned by recombineering from a clone of RPCI-23 C57BL/6J mouse genomic DNA library.

Please make sure the plasmid DNA preparation is pure by gel electrophoresis before linearizing for an electroporation.

Please note there are a few ambiguous bases in Addgene’s NGS result. This is due to repetitive regions of G-T and A-C in the 5' and 3' homology regions. Depositor confirms these bases do not affect plasmid function.