pET28a-HaloTag7SS-DogTag
(Plasmid #171775)

• Purpose: Expresses in bacterial cytoplasm HaloTag7 with DogTag inserted between D139 and E140, as well as C61S and C261S in HaloTag7 to block disulfide bond formation
• Depositing Lab: Mark Howarth
• Publication: Keeble et al Cell Chem Biol. 2021 Jul 28. pii: S2451-9456(21)00315-9. doi: 10.1016/j.chembiol.2021.07.005.

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 6401
• Vector type: Bacterial Expression

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: HaloTag7SS-DogTag
• Species: Synthetic
• Insert Size (bp): 1032
• Mutation: DogTag inserted in HaloTag7 between residues D139 and E140 and C61S and C261S mutations in HaloTag7 to block disulfide bond formation
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7F
• 3′ sequencing primer: T7R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • HaloTag Vector Limited Use Label License
• Industry Terms:
  • Not Available to Industry