p361_DG
(Plasmid #234683)

• Purpose: (Empty Backbone) Golden gate domesticated pET28b with His6-3C-3XFLAG-SUMO and Thrombin-HiBiT
• Depositing Lab: Matthew Wohlever
• Publication: Gaur et al Protein Sci. 2025 Apr;34(4):e70106. doi: 10.1002/pro.70106.

Backbone

• Vector backbone: pET28b
• Vector type: Bacterial Expression
• Tags / Fusion Proteins:
  • His6-3C-3XFLAG-SUMO (N terminal on backbone)
  • Thrombin-HiBiT (C terminal on backbone)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: None
• Mutation: No
• Tags / Fusion Proteins:
  • His6-3C-3XFLAG-SUMO (N terminal on backbone)
  • Thrombin-HiBiT (C terminal on backbone)

Terms and Licenses

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

Depositor Comments

The construct features an N-terminal His6 tag followed by a 3C protease cleavage site, a 3_ FLAG tag, and a SUMO tag. The C-terminus includes a Thrombin cleavage site followed by a HiBiT tag. BsaI recognition sites flank mScarlet3 to enable easy visualization of successful cloning through loss of red color. 5' Cloning Site: BsaI (destroyed), 3' Cloning Site: BsaI (destroyed).