pcDNA3-Lyso-AktAR2
(Plasmid #64933)

• Purpose: Lysosome-targeted FRET biosensor for monitoring Akt activity; targeted to cytosolic face of lysosomal membrane.
• Depositing Lab: Jin Zhang
• Publication: Zhou et al Cell Rep. 2015 Mar 11. pii: S2211-1247(15)00177-1. doi: 10.1016/j.celrep.2015.02.031.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5446
• Total vector size (bp): 8697
• 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: LAMP1-AktAR2
• Species: H. sapiens (human), S. cerevisiae (budding yeast), Synthetic
• Insert Size (bp): 3282
• Entrez Gene: LAMP1 (a.k.a. CD107a, LAMPA, LGP120)
• Promoter: CMV
• Tags / Fusion Proteins:
  • Cerulean3 (N terminal on insert)
  • cpVenus[E172] (C terminal on insert)
  • Full-length LAMP1 (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: T7 forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-Lyso-AktAR2

Terms and Licenses

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