pcDNA3-mCherry-TSC2(AA)-NLS
(Plasmid #192443)

• Purpose: Encodes mCherry-tagged TSC2 isoform 5 with S939A, T1439A mutations to block Akt phosphorylation; targeted to the nucleus.
• Depositing Lab: Jin Zhang
• Publication: Zhong et al Cell Chem Biol. 2022 Jun 16;29(6):1037-1045.e4. doi: 10.1016/j.chembiol.2022.02.006. Epub 2022 Mar 15.

Backbone

• Vector backbone: pcDNA3
• Backbone size w/o insert (bp): 5472
• Total vector size (bp): 11589
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: mCherry-TSC2(S939A,T1439A)-NLS
• Species: H. sapiens (human)
• Insert Size (bp): 6117
• Mutation: TSC2 Ser 939 and Thr 1439 (equivalent to 1462 in isoform 1) mutated to Ala.
• Entrez Gene: TSC2 (a.k.a. LAM, PPP1R160, TSC4)
• Promoter: CMV
• Tags / Fusion Proteins:
  • 6xHIS - T7 tag (gene 10 leader) - Xpress (TM) tag (N terminal on backbone)
  • mCherry (N terminal on insert)
  • Nuclear localization signal (NLS) (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-mCherry-TSC2(AA)-NLS.gbk
• A portion of this plasmid was derived from a plasmid made by: TSC2-AA cDNA was provided by Brendan Manning (Addgene Plasmid #14131)

Terms and Licenses

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