Double UP mNeon to mScarlet Rac1-V12
(Plasmid #125136)

• Purpose: Natively produces mNeonGreen, in the presence of Cre will be recombined to produce mScarlet and an untagged Rac1-V12 (Constitutively Active).
• Depositing Lab: Erik Dent
• Publication: Taylor et al Front Mol Neurosci. 2020 May 20;13:82. doi: 10.3389/fnmol.2020.00082. eCollection 2020.

Backbone

• Vector backbone: pCax
• Backbone size w/o insert (bp): 6734
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: mNeon/mScarletP2A Rac1-V12
• Alt name: Rac1-V12
• Alt name: 5879
• Mutation: Removed Not1 site in mScarlet. 12th AA in Rac1 changed to V
• Promoter: CAG

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CAGCTCCTGGGCAACGTGC
• 3′ sequencing primer: m13R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • mNeonGreen Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Intent is to introduce low doses of pCAG Cre to recombine a subset of cells, such that mNeon can act as an internal control to mScarlet for use in in utero electroporation or other related techniques. Rac1-V12 is expressed following a P2A site, so that Cre+ cells will be both mScarlet positive and also contain an untagged Rac1-V12.

Note: Addgene NGS is unable to fully resolve the CAG promoter sequence. Please refer to the depositor's sequence for this section of the plasmid.