pCMV-104
(Plasmid #79842)

• Purpose: Expresses NLS-PpsR2-VP16 under CMV promoter
• Depositing Lab: Vladislav Verkhusha
• Publication: Kaberniuk et al Nat Methods. 2016 May 9. doi: 10.1038/nmeth.3864.

Backbone

• Vector backbone: pEGFP-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4000
• Total vector size (bp): 5900
• 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-PpsR2-VP16
• Species: Rhodopseudomonas palustris
• Insert Size (bp): 1900
• Mutation: RpPpsR2 cysteine 439 changed to serine
• GenBank ID: KX063612 CAE26978.1
• Promoter: CMV
• Tags / Fusion Proteins:
  • VP16 (C terminal on insert)
  • Nuclear localization signal of nuclear cap-binding protein subunit 1 (NLS) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AsisI (not destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: ggtaggcgtgtacggtgggag

Resource Information

• Supplemental Documents:
  • pCMV-104
• A portion of this plasmid was derived from a plasmid made by: A RpPpsR2 gene of R. palustris (NCBI Gene rpa1536) (accession code CAE26978) was provided by M. Papiz (Liverpool University, UK) and T. Beatty (University of British Columbia, Canada).

Terms and Licenses

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

Depositor Comments

New paper providing detailed protocol for using this plasmid- https://www.nature.com/articles/nprot.2018.022