pKK-BiFC-Venus
(Plasmid #105804)

• Purpose: (Empty Backbone) Concomitant expression of two proteins in fusion with fragments of Venus protein. Protein interactions studies by bimolecular fluorescence complementation (BiFC), including BiFC-FRET assays.
• Depositing Lab: Andrzej Dziembowski
• Publication: Szczesny et al PLoS One. 2018 Mar 28;13(3):e0194887. doi: 10.1371/journal.pone.0194887. eCollection 2018.

Backbone

• Vector backbone: BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen)
• Backbone manufacturer: Sammarco & Grabczyk (PMID: 16212928)
• Vector type: Mammalian Expression ; Flp-In competent
• Selectable markers: Hygromycin
• Tag / Fusion Protein:
  • VN173 (I152L); VC155 (C terminal on backbone)

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: None
• Tag / Fusion Protein:
  • VN173 (I152L); VC155 (C terminal on backbone)

Resource Information

• Supplemental Documents:
  • pKK-BiFC-Venus.txt
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Addgene has found the VN173 mutation (I152L) is not present in this plasmid.

This vector is a member of the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101) pKK-BiFC-Venus vector encodes fragments of Venus (VN173, VC155) that become fluorescent when they reassemble into full fluorescent protein. Thus, it is useful for protein-protein interactions studies involving bimolecular fluorescence complementation approach (BiFC) (PMID: 16454041, 18846096). Here, an improved version of VN173 fragment is used (I152L) to inhibit self-assembly (PMID:21091444). Venus fragments coding sequences were cloned from Addgene ID: 22010 and 22011 and VN173 was subjected to site-directed mutagenesis to introduce mutation I152L.