Syk tSH2 R44A in pET-30a(+)
(Plasmid #172856)

• Purpose: express murine Syk tandem SH2 domains (Ser 8 to Gln 264), with R44A mutation and a TEV-cleavable His-tag. For use in E.coli strain Rosetta 2 (DE3).
• Depositing Lab: Carol Post
• Publication: mutants for salt bridges found in syk md simulations (unpublished)

Backbone

• Vector backbone: pET-30a(+)
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5422
• Total vector size (bp): 6081
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Provided by Addgene in DH5alpha. Use Rosetta 2 (DE3) for expression studies.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: murine Syk tandem SH2 domains (Ser 8 to Gln 264), with R44A mutation and a TEV-cleavable His-tag
• Alt name: Spleen tyrosine kinase
• Species: M. musculus (mouse)
• Insert Size (bp): 852
• Mutation: R44A
• GenBank ID: NP_035648
• Entrez Gene: Syk (a.k.a. Sykb)
• Promoter: T7
• Tag / Fusion Protein:
  • TEV-cleavable His-tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (unknown if destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Terms and Licenses

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

Depositor Comments

encoding the following protein sequence: MGSSHHHHHHDYDIPTTENLYFQSANHLTYFFGNITREEAEDYLVQGGMTDGLYLLRQSANYLGGFALSVAHNRKAHHYTIERELNGTYAISGGRAHASPADLCHYHSQEPDGLICLLKKPFNRPPGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGNISRDESEQTVLIGSKTNGKFLIRARDNSGSYALCLLHEGKVLHYRIDRDKTGKLSIPEGKKFDTLWQLVEHYSYKPDGLLRVLTVPCQKIGAQ–