pEN_TT 3xFLAG-MAVS-Gln93,Ala271
(Plasmid #158632)

• Purpose: Gateway entry vector for an inducible 3xFLAG-tagged MAVS mutant
• Depositing Lab: Kevin Janes
• Publication: Lopacinski et al Cell Syst. 2021 Apr 21;12(4):304-323.e13. doi: 10.1016/j.cels.2021.02.004. Epub 2021 Mar 18.

Backbone

• Vector backbone: pEN_TTmiR2c_3xFLAG
• Backbone manufacturer: Addgene #83274
• Vector type: Gateway entry vector

Growth in Bacteria

• Bacterial Resistance(s): Gentamicin, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: MAVS
• Alt name: IPS-1
• Alt name: VISA
• Species: H. sapiens (human)
• Mutation: E93Q/Q271A
• Entrez Gene: MAVS (a.k.a. CARDIF, IPS-1, IPS1, VISA)
• Tag / Fusion Protein:
  • 3XFLAG (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: MfeI (not destroyed)

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The plasmid differs from the MAVS reference sequence at 2 positions (Q198K and S409F). This plamid has an additional Q271A mutation, and amino acid 93 (glutamate, E) was mutated back to the reference (glutamine, Q). Gene point mutations were performed using the QuikChange Kit (Agilent Cat#200517) two times on the pEN_TT 3XFLAG-MAVS plasmid: one mutagenesis to yield pEN_TT 3XFLAG-MAVS-Q271A and a second mutagenesis to yield the double mutant. The clones displayed consistent off-target mutations at one of two points. Two clones with different off-target mutations were digested and the good pieces ligated together so to yield the final sequence.