pDN80
(Plasmid #61349)

• Purpose: (Empty Backbone) LINuS-flex: modular LINuS construct enabling introduction of any protein encoding sequence, NES and NLS. The sequence of interest replaces the LexA DBD-MBP-VP64 fragment.
• Depositing Labs: Barbara Di Ventura, Roland Eils
• Publication: Niopek et al Nat Commun. 2014 Jul 14;5:4404. doi: 10.1038/ncomms5404.

Backbone

• Vector backbone: pmCherry-N1
• Backbone manufacturer: Clontech
• Vector type: Mammalian Expression
• Promoter: CMV
• Tags / Fusion Proteins:
  • mCherry (C terminal on backbone)
  • AsLOV2 wt (C terminal on backbone)

Growth in Bacteria

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

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: mCherry-R

Resource Information

• Supplemental Documents:
  • pDN80 genbank

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

LINuS-flex was designed to contain a spacer sequence flanked by BsmBI restriction sites between the VP64 transactivation domain and mCherry enabling scarless introduction of any NES sequence by standard oligo cloning.
A second spacer sequence flanked by BsaI restriction sites is located behind the truncated AsLOV2 domain and enables scarless introduction of any NLS sequence by standard oligo cloning.
Unique BamHI and SpeI sites further enable the exchange of the LexA DBD-MBP-VP64 fragment by any sequence of interest.