pcDNA3.1hSIRT3_H248Y_HA
(Plasmid #24492)

• Depositing Lab: Eric Verdin
• Publication: Hirschey et al Nature. 2010 Mar 4. 464(7285):121-5.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• 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: Unknown

Gene/Insert

• Gene/Insert name: Human Sirtuin 3 catalytically inactive mutant
• Alt name: SIRT3
• Alt name: SIRT3-H248Y
• Species: H. sapiens (human)
• Insert Size (bp): 1000
• Mutation: H248Y
• GenBank ID: NM_012239
• Entrez Gene: SIRT3 (a.k.a. SIR2L3)
• Tag / Fusion Protein:
  • HA (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

SIRT3 contains a mutation at amino acid Q32H. This mutation is present in the first 100 amino acids of SIRT3 which serve as its mitochondrial import signal. A peptide corresponding to the first 100 aa of SIRT3 is normally cleaved once the protein has been imported into the mitochondria and the active form of SIRT3 in the mitochondrial matrix therefore lacks this part of the protein. Therefore the mutation should not modify SIRT3 activity.