pGTM-Sumo2-WN-NS2_3_TEE
(Plasmid #254047)

• Purpose: Expresses West Nile Virus NS3 and an NS2 fragment on a polyistronic message for E. coli expression
• Depositing Lab: Gaetano Montelione
• Publication: A PA CLAN PROTEASE CROSS-INHIBITION SCREEN (unpublished)

Backbone

• Vector backbone: pET15Sumo2_NESG
• Backbone manufacturer: Novagen/Northeast Structural Genomics Consortium
• Backbone size w/o insert (bp): 5680
• Total vector size (bp): 6413
• Modifications to backbone: An N-terminal 6xHis tag fused to the Smt3 gene (Sumo) of Saccharomyces cerevisiae as a 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).
• 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 1

• Gene/Insert name: NS3
• Species: West Nile Virus
• Insert Size (bp): 558
• GenBank ID: AFP50427
• Promoter: T7 lac
• Tag / Fusion Protein:
  • 6xHis, Sumo (N terminal on backbone)

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: NS2
• Alt name: AFP50427
• Species: West Nile Virus
• Insert Size (bp): 150
• Promoter: Expressed as the second ORF in a polycistronic message

Cloning Information for Gene/Insert 2

• 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