pAAV-CMVd1-NLS-GAL4-iLight2-VP16
(Plasmid #219550)

• Purpose: NLS-Gal4(1-65)-iLight2-VP16 construc expression in mammalian cells. pAAV vector.
• Depositing Lab: Vladislav Verkhusha
• Publication: Baloban et al Protein Sci. 2024 May;33(5):e4993. doi: 10.1002/pro.4993.

Backbone

• Vector backbone: pAAV-CW3SL-EGFP
• Backbone manufacturer: Addgene #61464
• Backbone size w/o insert (bp): 3515
• 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: NLS-Gal4(1-65)-iLight2-VP16
• Species: Synthetic
• Insert Size (bp): 2346
• Mutation: The (SAGG)x4 linker between GAL4(1-65) and the PCM of IsPadC was replaced with a (SAGG)x1 linker.
• GenBank ID: MW890755.1 (part of the insert)
• Promoter: CMVd1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: pBR322ori-F
• 3′ sequencing primer: WPRE-R

Terms and Licenses

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

Depositor Comments

To make pAAV-CMVd1-NLS-GAL4(1-65)-iLight2-VP16, NLS-GAL4-(SAGG)x4 was first replaced with NLS-GAL4-SAGG in the partiall modified pAAV-CaMKII-GAL4-iLight-VP16 (Addgene #170274) using the NheI and EcoR restriction sites, followed by the introduction of an additional NheI site upstream of the CaMKII promoter, and finally, the CaMKII promoter was swapped with CMVd1 using NheI restriction sites.