pB6-Csrp2-T2A-sfGFP-iCre-ERT2-FNF-DTA
(Plasmid #140270)

• Purpose: B6J Csrp2-T2A-sfGFP-iCre-ERT2 allele targeting vector, low copy number
• Depositing Lab: Mario Capecchi
• Publication: Miscellaneous gene targeting vector plasmids (unpublished)

Backbone

• Vector backbone: pDTA-4
• Backbone manufacturer: Hyung-song Nam, Capecchi laboratory
• Vector type: Mammalian Expression, Mouse Targeting, Cre/Lox
• 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: Csrp2
• Species: M. musculus (mouse)
• Entrez Gene: Csrp2 (a.k.a. AW551867, Crp2, SmLim)
• Tag / Fusion Protein:
  • C-terminal fusion of T2A-sfGFP-iCre-ERT2 (C terminal on insert)

Resource Information

• Supplemental Documents:
  • pCsrp2-T2A-GFPcreER-DTA4.gb
• A portion of this plasmid was derived from a plasmid made by: sfGFP is from Sandia Biotech. iCre is from Rolf Sprengel. ERT2 is from Pierre Chambon. FRT-Neo-FRT is from pFNF (#22687).

Terms and Licenses

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

Depositor Comments

Linker between sfGFP and iCre-ERT2 is a mutated P2A sequence where 2 conserved amino acids are changed to Alanines. This mutated sequence does not skip.

The sfGFP fluorescence from the sfGFP-iCre-ERT2 is dim, as described in Nam and Capecchi, 2020 (https://doi.org/10.1186/s13064-020-00139-5) and a Figshare page (https://doi.org/10.6084/m9.figshare.13218437.v35). Thus, the sfGFP fluorescence should be detectable only in cells in which the marker gene expression level is high.

Homology arms subcloned by recombineering from a clone of RPCI-24 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 T-A regions. Depositor confirms these bases do not affect plasmid function.