Double UP mClover3 to mScarlet
(Plasmid #120262)

• Purpose: Natively produces mClover3, in the presence of Cre will be recombined to produce mScarlet.
• 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
• 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: mClover3/mScarletP2A-mcs
• Mutation: Removed Not1 site in mScarlet
• Promoter: CAG

Cloning Information

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

Resource Information

• Supplemental Documents:
  • double-up-mclover3-to-scarlet.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• 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 mClover3 can act as an internal control to mScarlet for use in in utero electroporation or other related techniques. Genetic overexpression can be easily attached following the mScarlet through use of a mcs after the P2A.

The associated publication utilized mNeon instead of sfGFP/mClover3, but mNeon has is exceedingly difficult to share. Therefore, these two plasmids (120261 and 120262) are made available, using sfGFP and mClover3 instead of mNeon. These plasmids have been tested, and are equivalent to the mNeon variant.

Please 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.