pN-PURO-PTP
(Plasmid #51711)

• Purpose: (Empty Backbone) contains an N-terminal tandem affinity purification tag (relevant to any system) within a construct designed for genome integration into trypanosomes.
• Depositing Lab: Arthur Gunzl
• Publication: Schimanski et al Eukaryot Cell. 2005 Nov;4(11):1942-50.

Backbone

• Vector backbone: pC-PTP-NEO (Addgene plasmid #51710)
• Modifications to backbone: The resistance marker cassette was modified by replacing NEO-R by the coding region of the puromycin N-acetyltransferase gene (PURO-R).
• Vector type: T. brucei genome integration vectors
• Selectable markers: Puromycin
• Tag / Fusion Protein:
  • PTP (ProtC-TEV-ProtA) (N 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: M13-F20
• 3′ sequencing primer: M13R

Terms and Licenses

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

Depositor Comments

The PTP tag is a derivative of the original TAP tag and has CBP replaced by ProtC.

The PTP cassette is composed of 434 bp of 5′ flank from the TbRPA2 gene, encoding the second-largest subunit of RNA polymerase I, a translation initiation codon, the PTP tag coding sequence, and unique restriction sites for fusing target sequences to the tag.