pGTM_AviHTSumoTOEET_NESG-HRV2-3CLpro
(Plasmid #254122)

• Purpose: Expresses HRV2 3CLpro in E. coli with fused Avi-tag, 6xHis and Sumo SET tag
• Depositing Lab: Gaetano Montelione
• Publication: A PA CLAN PROTEASE CROSS-INHIBITION SCREEN (unpublished)

Backbone

• Vector backbone: pET15AviHTSumoTOEET
• Backbone manufacturer: Novagen/Northeast Structural Genomics Consortium
• Backbone size w/o insert (bp): 5727
• Total vector size (bp): 6273
• Modifications to backbone: An N-terminal Avi-tag is in frame with a 6xHis tag followed by a TEV recognition site and fused to an expression optimized Smt3 gene (Sumo) of Saccharomyces cerevisiae as an expression/purification/solubility enhancement cassette. BsaI sites are included in the cloning polylinker for scarless cloning and the production of a native, mature protein after cleavage from the Sumo fusion by Sumo protease (Ulp1). This vector uses the Transcript Optimized Expression Enhancement Technology described in Patent #10,385,350.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Utilize BL21DE3 for protein expression.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: 3C protease
• Species: Rhinovirus Type 2
• Insert Size (bp): 546
• GenBank ID:
• Promoter: T7
• Tag / Fusion Protein:
  • Avi-tag, 6xHis, TEV site, Sumo (N terminal on backbone)

Cloning Information

• Cloning method: Gene Synthesis
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Genscript synthesized with codon optimization for E. coli

Terms and Licenses

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

Depositor Comments

Related to PDB: 1CQQ