pCMVd1-NLS-GAL4-iLight2-VP16-T2A-mTagBFP2
(Plasmid #219548)

• Purpose: NLS-Gal4(1-65)-iLight2-VP16-T2A-mTagBFP2 construct expression in mammalian cells.
• Depositing Lab: Vladislav Verkhusha
• Publication: Baloban et al Protein Sci. 2024 May;33(5):e4993. doi: 10.1002/pro.4993.

Backbone

• Vector backbone: pQP-Select
• Backbone manufacturer: Originally derived from pEGFP-N1 backbone after several modifications in our lab.
• Backbone size w/o insert (bp): 3427
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: NLS-Gal4(1-65)-iLight2-VP16-T2A-mTagBFP2
• Species: Synthetic
• Insert Size (bp): 3118
• 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: BamHI (not destroyed)
• 5′ sequencing primer: pBR322ori-F
• 3′ sequencing primer: F1ori-F

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

To make pCMVd1-NLS-GAL4-iLight2-VP16-T2A-mTagBFP2, NLS-GAL4-Linker fragment was amplified and swapped with NLS-GAL4-4x(SAGG) fragment in pCMVd1-NLS-GAL4-iLight-VP16-T2A-mTagBFP2 plasmid (Addgene #170273) using NheI and EcoRI restriction sites.