pCAGGS-PHOSPHO1-TEV-Twin-Strep
(Plasmid #222015)

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• Publication: PHOSPHO2 is PA phosphatase and PHOSPHO1 is PLC (unpublished)

Backbone

• Vector backbone: pCAGGS-N-TEV-Twin-Strep
• Backbone manufacturer: Chiaki Murakami
• Backbone size w/o insert (bp): 4871
• Total vector size (bp): 5672
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: PHOSPHO1
• Alt name: phosphoethanolamine/phosphocholine phosphatase 1
• Species: H. sapiens (human)
• Insert Size (bp): 801
• Mutation: stop codon (TGA) is deleted
• GenBank ID: NM_178500.4
• Entrez Gene: PHOSPHO1
• Entrez Gene: PHOSPHO1
• Promoter: chicken β-actin 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: TTCGGCTTCTGGCGTGTGACC
• 3′ sequencing primer: TATTAGCCAGAAGTCAGATGCTCAAGG

Terms and Licenses

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

Depositor Comments

The insert can be removed using the restriction enzyme, XhoI.