pET-15b-TEV-C9R
(Plasmid #118754)

• Purpose: TEV-C9R protease expression vector with enhanced expression, solubility and yield without affecting TEV's protease activity
• Depositing Lab: Yutaka Kuroda
• Publication: Nautiyal et al N Biotechnol. 2018 May 25;42:77-84. doi: 10.1016/j.nbt.2018.02.006. Epub 2018 Feb 12.

Backbone

• Vector backbone: pET-15b
• Backbone size w/o insert (bp): 5691
• Total vector size (bp): 6512
• Modifications to backbone: No modification
• 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: Tobacco Etch Virus protease
• Alt name: TEV-C9R
• Species: Synthetic
• Insert Size (bp): 768
• Entrez Gene: NEWENTRY
• Promoter: T7
• Tags / Fusion Proteins:
  • C9R (C terminal on insert)
  • His tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nde1 (not destroyed)
• 3′ cloning site: BamH1 (not destroyed)
• 5′ sequencing primer: taatacgactcactatagg
• 3′ sequencing primer: ccgctgagcaataactagc

Terms and Licenses

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

Depositor Comments

Japanese patent pending (2017-247019)
Laboratory URL: http://www.tuat.ac.jp/~ykuroda/y_index_e.html

A frameshift in the lacI sequence was detected by Addgene NGS, but it probably does not affect the expression level as the same plasmid was used for expression check in the paper.