pGEX-PHOSPHO2-TEV-Twin-Strep
(Plasmid #222017)

• Purpose: Express human PHOSPHO2 protein (C-terminal Tobacco Etch Virus (TEV) protease cleavable Twin-Strep-fusion protein (ENLYFQGS-WSHPQFEK-(GGGS)2-GGSA-WSHPQFEK)) in E. coli
• Depositing Lab: Chiaki Murakami
• Publication: PHOSPHO2 is PA phosphatase and PHOSPHO1 is PLC (unpublished)

Backbone

• Vector backbone: pGEX-N-TEV-Twin-Strep
• Backbone manufacturer: Chiaki Murakami
• Backbone size w/o insert (bp): 3679
• Modifications to backbone: The TEV cleavable site (ENLYFQG) and Twin-strep tag were inserted.
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: PHOSPHO2
• Alt name: phosphatase, orphan 2
• Species: H. sapiens (human)
• Insert Size (bp): 723
• Mutation: deleted stop codon (TAA)
• GenBank ID: NM_001008489.4
• Entrez Gene: PHOSPHO2
• Promoter: lac promoter
• Tags / Fusion Proteins:
  • TEV cleavable site (C terminal on backbone)
  • Twin-Strep-tag (C terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CATTAGGCACCCCAGGCTTTACACTTTATG
• 3′ sequencing primer: GCTTACAGACAAGCTGTGACCGTC

Terms and Licenses

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

Depositor Comments

Expression Conditions:

OD600: approximately 0.5~0.8
IPTG: approximately 0.5 mM
37°C, 3 h