TOPO mMecp2-NLucTom
(Plasmid #159282)

• Purpose: To tag endogenously and c-terminally mouse Mecp2 with two reporters: NanoLuciferase and TdTomato. It contains 2 homology arms from a castaneus background surrounding the STOP codon.
• Depositing Lab: Joost Gribnau
• Publication: Mira-Bontenbal et al Stem Cell Reports. 2022 Jan 29. pii: S2213-6711(22)00052-2. doi: 10.1016/j.stemcr.2022.01.008.

Backbone

• Vector backbone: TOPO
• Backbone size w/o insert (bp): 3424
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Mecp2
• Species: M. musculus (mouse)
• Insert Size (bp): 3157
• Mutation: Nluciferase-TdTomato fusion
• Entrez Gene: Mecp2 (a.k.a. 1500041B07Rik, D630021H01Rik, Mbd5, WBP10)
• Tag / Fusion Protein:
  • Nluciferase-P2A-TdTomato (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: SP6

Resource Information

• Supplemental Documents:
  • TOPO mMecp2-NLucTom Addgene.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • NanoLuc Luciferase Limited Use Label License
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Both homology arms are from a castaneus background. This plasmid was used to target the C-terminus of mouse Mecp2 to fuse it in frame with a NanoLuciferase reporter and an additional TdTomato. There is a P2A signal between Nluc and Tomato leading to Tomato being translated as an independent protein.