pCAG-sfGFP-iCre-ERT2
(Plasmid #125749)

• Purpose: sfGFP and iCreERT2 fused by unskippable linker
• Depositing Lab: Mario Capecchi
• Publication: Nam et al Neural Dev. 2020 Mar 17;15(1):3. doi: 10.1186/s13064-020-00139-5.

Backbone

• Vector backbone: Custom
• Vector type: Mammalian Expression, Cre/Lox

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: sfGFP-iCre-ERT2
• Species: Synthetic

Resource Information

• Supplemental Documents:
  • CAG sfGFP-linker-iCreERT2.gb
• A portion of this plasmid was derived from a plasmid made by: CAG promoter is from pCAGGS obtained from BCCM/LMBP. sfGFP is from Sandia Biotech. iCre is from Rolf Sprengel. ERT2 is from Pierre Chambon. WPRE is from Miguel Ramalho-Santos. bGH pA is from Neal Copeland. Backbone is from Stratagene.

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) In short, it is not visible to the eyes, but can be detected with flow cytometry.

Similar to the situation with some other plasmids in this collection, the missing four nucleotides at the 5' end of the WPRE renders it unrecognizable to SnapGene, but the expression vector is functional.