pNLS-2xBFP(4C)
(Plasmid #176151)

• Purpose: Encodes NLS-2xBFP(4C), which is a tandem BFP with an N-terminal SV40 NLS. The protein has 4 solvent accessible cysteines for labeling with maleimide dyes.
• Depositing Lab: Siegfried Musser
• Publication: Chowdhury et al Nat Cell Biol. 2022 Jan;24(1):112-122. doi: 10.1038/s41556-021-00815-6. Epub 2022 Jan 10.

Backbone

• Vector backbone: pTrcHisB
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4404
• Total vector size (bp): 5780
• Vector type: Bacterial Expression

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: NLS-2xBFP(4C)
• Species: Aequorea victoria
• Insert Size (bp): 1509
• Mutation: Compared with EGFP, the two BFP domains have the following mutations: L64F, Y66H, Q80R, V160A, S174C, and L230H
• GenBank ID: AFA52654.1
• Promoter: Ptrc
• Tags / Fusion Proteins:
  • SV40 nuclear localization sequence (NLS) (N terminal on insert)
  • 6xHis tag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: rrnG antiterminator
• 3′ sequencing primer: rrnB T1 terminator

Resource Information

• Supplemental Documents:
  • pNLS-2xBFP(4C).gbk

Terms and Licenses

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