pQE30-butelase1
(Plasmid #115430)

• Purpose: Expresses the macrocyclizing asparaginyl endopeptidase butelase 1 in E. coli
• Depositing Lab: Joshua Mylne
• Publication: James et al Plant J. 2019 Feb 21. doi: 10.1111/tpj.14293.

Backbone

• Vector backbone: pQE30
• Backbone manufacturer: QIAGEN
• Backbone size w/o insert (bp): 3450
• Total vector size (bp): 4831

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: butelase 1
• Alt name: CtAEP1
• Species: Synthetic
• Insert Size (bp): 1400
• Promoter: T5
• Tag / Fusion Protein:
  • 6-His (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: CCCGAAAAGTGCCACCTG
• 3′ sequencing primer: GTTCTGAGGTCATTACTGG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Is a synthetic gene, codon optimised for E. coli made upon request by GENEART (14AB5UDP)

Terms and Licenses

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

Depositor Comments

Expression and purification details in bioRxiv preprint “The macrocyclizing protease butelase 1 remains auto-catalytic and reveals the structural basis for ligase activity” by Amy M James, Joel Haywood, Julie Leroux, Katarzyna Ignasiak, Alysha G Elliott, Jason W Schmidberger, Mark F Fisher, Samuel G Nonis, Ricarda Fenske, Charles S Bond, Joshua S Mylne
doi: https://doi.org/10.1101/380295 . Please visit https://www.biorxiv.org/content/early/2018/07/30/380295 for bioRxiv preprint.