pKAR2-Br512_Mut235
(Plasmid #179278)

• Purpose: Expresses Br512_Mut235 polymerase in E.coli
• Depositing Lab: Andrew Ellington
• Publication: Paik et al Biochemistry. 2021 Nov 11. doi: 10.1021/acs.biochem.1c00451.

Backbone

• Vector backbone: pKAR2
• Backbone manufacturer: In-house made
• Backbone size w/o insert (bp): 5187
• Total vector size (bp): 6218
• 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: Engineered Bst DNA Polymerase I Large Fragment
• Alt name: Br512g2.2, Br512_Mut235, Mut235
• Species: Geobacillus stearothermophilus
• Insert Size (bp): 1932
• GenBank ID: 4O0I_A
• Promoter: T7
• Tag / Fusion Protein:
  • 8x His tag; Villin Headpiece 47 amino acid (HP47) ; Linker (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (unknown if destroyed)
• 3′ cloning site: BsaI (unknown if destroyed)
• 5′ sequencing primer: CCTATAAAAATAGGCGTATCACGAGGC
• 3′ sequencing primer: ACCCGTTTAGAGGCCCCAAGG

Resource Information

• Supplemental Documents:
  • pKAR2_Br512_Mut235.gb

Terms and Licenses

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