MBP-TEVcs(ENLYFQ/G)-His6-uTEV1Δ(220-242)-R5
(Plasmid #135455)

• Purpose: Bacterial expression of uTEV1Δ protease
• Depositing Lab: Alice Ting
• Publication: Sanchez et al Nat Methods. 2019 Dec 9. pii: 10.1038/s41592-019-0665-7. doi: 10.1038/s41592-019-0665-7.

Backbone

• Vector backbone: pMal-C2
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 5479
• Total vector size (bp): 7357
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Use BL21(DE3)-RIL for protein expression. Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble uTEV1Δ protease. Ampicillin for MBP-TEVcs(ENLYFQ/G)-His6-uTEV1Δ(220-242)-R5; Chloramphenicol for pRIL.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: MBP-TEVcs(ENLYFQ/G)-His6-TEV1Δ(220-242)-R5
• Species: Synthetic
• Insert Size (bp): 1878
• Mutation: S219V mutation improves stability, S153N mutation improves catalytic activity. Truncated form of TEV protease (low affinity)
• Promoter: tac promoter
• Tag / Fusion Protein:
  • MBP, His6

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: N/A
• 3′ sequencing primer: M13-F20

Resource Information

• Supplemental Documents:
  • MBP-TEVcs(ENLYFQG)-His6-uTEV1Δ(220-242)-R5

Terms and Licenses

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