pT2ADW-lox-tdKeima-NES-lox-6011nls(ERK)-P2Av3-RGECO1
(Plasmid #162486)

• Purpose: For generating transgenic mice expressing ERK biosensor and RGECO1
• Depositing Lab: Michiyuki Matsuda
• Publication: Imanishi et al Am J Pathol. 2020 Oct 15. pii: S0002-9440(20)30459-4. doi: 10.1016/j.ajpath.2020.09.012.

Backbone

• Vector backbone: pT2ADW
• Backbone size w/o insert (bp): 7300
• Modifications to backbone: pT2ADW was constructed as follows: The D4Z4 insulator was inserted before the CAG promoter of pT2AL200R175 carrying Tol2 recombination sites. Then, the WPRE sequence of pCSII-EF was inserted before the poly A sequence.
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: ERK biosensor based on FRET, and RGECO1
• Species: Synthetic
• Insert Size (bp): 5000
• Promoter: CAG

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: NruI (not destroyed)
• 5′ sequencing primer: pCAGGS-5

Resource Information

• A portion of this plasmid was derived from a plasmid made by: cDNA coding RGECO1 was a gift from Dr. Robert Campbell (Addgene plasmid # 32444 ; http://n2t.net/addgene:32444 ; RRID:Addgene_32444)

Terms and Licenses

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