pET23a-H6-TEV-AncSZ
(Plasmid #214235)

• Purpose: Bacterial expression construct of AncSZ, an kinase domain engineered through ancestorial reconstruction of the kinase domains of both SYK and ZAP-70. For in vitro protein tyrosine phosphorylation
• Depositing Lab: Neel Shah
• Publication: Hobbs et al Protein Sci. 2022 Oct;31(10):e4411. doi: 10.1002/pro.4411.

Backbone

• Vector backbone: pET23a
• Backbone size w/o insert (bp): 3700
• Total vector size (bp): 4534
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Syk kinase
• Species: H. sapiens (human)
• Insert Size (bp): 834
• Mutation: Sequence gained through ancestral reconstruction of Syk and ZAP-70 kinase domains and codon optimized for E. coli expression
• Entrez Gene: SYK (a.k.a. IMD82, p72-Syk)
• Promoter: T7
• Tag / Fusion Protein:
  • 6xHis (N terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 promotor

Resource Information

• Supplemental Documents:
  • Shah_H7Y_4_pET-23a-H6-TEV-AncSZ-KD_pLann.gbk

Terms and Licenses

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

Depositor Comments

This protein needs to be co-expressed in E. coli BL21(DE3) cells with the YopH tyrosine phosphatase