pTet-On-TAP
(Plasmid #59017)

• Purpose: (Empty Backbone) Construct allows tetracycline inducible expression of a protein of interest fused to a tandem affinity purification tag (TAP) in toxoplasma gondii.
• Depositing Lab: David Sibley
• Publication: Etheridge et al Cell Host Microbe. 2014 May 14;15(5):537-50. doi: 10.1016/j.chom.2014.04.002.

Backbone

• Vector backbone: pTet07Sag1mycGFP
• Backbone manufacturer: Dominique Soldati Laboratory
• Backbone size (bp): 4531
• Modifications to backbone: Digested with Kpn1/PacI followed by insertion of a Tet regulatable RPS13 promoter (KpnI/MfeI) and multiple cloning site and TAP tag (MfeI/PacI). Performed a 3 fragment ligation of the pieces into the Tet-On-TAP vector.
• Vector type: Toxoplasma Expression
• Promoter: RPS13 with Tet Operators Inserted
• Selectable markers: Phleomycin
• Tag / Fusion Protein:
  • Calmodulin Binding Peptide fused with a TEV Cleavage site to Protein A-tag (C terminal on backbone)

Growth in Bacteria

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

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: M13F
• 3′ sequencing primer: M13R

Resource Information

• Supplemental Documents:
  • Gene Bank Vector Map

Terms and Licenses

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