pGEX6P1‐Afu‐rnhB D101N
(Plasmid #108695)

• Purpose: For expression in E. coli, and affinity purification of N-terminally GST-tagged Archaeoglobus fulgidus RNase HII with D101N catalytic site mutation
• Depositing Labs: Andrew Jackson, Martin Reijns
• Publication: Bubeck et al Nucleic Acids Res. 2011 May;39(9):3652-66. doi: 10.1093/nar/gkq980. Epub 2011 Jan 17.

Backbone

• Vector backbone: pGEX6P1
• Backbone manufacturer: Amersham
• Backbone size w/o insert (bp): 4984
• Total vector size (bp): 5606
• Modifications to backbone: SacI site added between between rnhB stop codon and NotI site
• 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: rnhB
• Species: Archaeoglobus fulgidus
• Insert Size (bp): 618
• Mutation: D101N catalytic site mutation
• Promoter: tac
• Tag / Fusion Protein:
  • GST (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: GGGCTGGCAAGCCACGTTTGGTG
• 3′ sequencing primer: CCGGGAGCTGCATGTGTCAGAGG

Resource Information

• Supplemental Documents:
  • pGEX6P1-Afu-rnhB-D101N.gb

Terms and Licenses

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