pET28a-ybbr-XMod-Doc (BMA-KO, S199AzF)-HIS
(Plasmid #153444)

• Purpose: E. coli expression (amber suppression) of Rc. XDocB binding mode A knock-out mutant with serine at position 199 replaced with amber codon.
• Depositing Lab: Michael Nash
• Publication: Liu et al Nat Commun. 2020 Aug 28;11(1):4321. doi: 10.1038/s41467-020-18063-x.

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 5219
• Total vector size (bp): 5966
• Vector type: Bacterial Expression

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: Rc.XDocB
• Alt name: Ruminococcus champanellensis XModule-dockerin B
• Species: Ruminococcus champanellensis
• Insert Size (bp): 747
• Mutation: Serine 199 was mutated to amber codon (TAG). Arginine 191 and leucine 195 were mutated to alanine and glutamic acid, respectively
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • ybbr tag (N terminal on insert)
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 forward primer
• 3′ sequencing primer: T7 reverse primer

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2020.01.21.913590v1 for bioRxiv preprint.