pAAV2-CAG-3xNLS-vsfGFP-0
(Plasmid #191097)

• Purpose: To express a bright green nanobody FP to label eucaryotic cell nuclei. To be used in tissue clearing methods and other fluorescent microscopy methods
• Depositing Labs: Murray Blackmore, Pantelis Tsoulfas
• Publication: Wang et al Elife. 2022 Jul 15;11. pii: 76254. doi: 10.7554/eLife.76254.

Backbone

• Vector backbone: pAAV2-CAG
• Backbone manufacturer: Original was from the Boyden laboratory
• Backbone size w/o insert (bp): 4714
• Total vector size (bp): 5860
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 3xNLS-vsfGFP-0
• Species: Synthetic
• Insert Size (bp): 1149
• Mutation: Optimized to Human codon usage
• Promoter: CMV and CAG
• Tag / Fusion Protein:
  • Green fluorescent protein bound to enhancer nanobody [Camelus dromedarius] Sequence ID: 3K1K_C (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (destroyed during cloning)
• 3′ cloning site: EcoRV (destroyed during cloning)
• 5′ sequencing primer: pCAGSS-5
• 3′ sequencing primer: WPRE-R

Terms and Licenses

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