pEntry Tshld-GFP
(Plasmid #53211)

• Purpose: pEntry Tshld-GFP is a product of BP recombination reaction between attB flanked Tshld system-encoded PCR product and pDonor 201 plasmid.
• Depositing Lab: Matthew Pratt
• Publication: Lin et al Chembiochem. 2014 Mar 11. doi: 10.1002/cbic.201400006.

Backbone

• Vector backbone: pDONR 201
• Backbone manufacturer: invitrogen
• Backbone size w/o insert (bp): 4470
• Total vector size (bp): 4073
• Modifications to backbone: Note that the insert was introduced by BP recombination.

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Tshld-GFP
• Alt name: FLAG tag-FRB-UbN(I13G)-T2A-FKBP*-UbC-HA tag-GFP
• Insert Size (bp): 1809
• Promoter: none
• Tags / Fusion Proteins:
  • Flag (N terminal on insert)
  • HA (C terminal on insert)
  • EGFP (C terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: TCGCGTTAACGCTAGCATGGATCTC
• 3′ sequencing primer: GTAACATCAGAGATTTTGAGACAC

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The Tshld system allows users to fine-tune the concentration of a protein of interest in mammalian cells by titrating the concentration of rapamycin or FK506. Unlike other post-translational techniques, the Tshld system releases its protein of interest in its native form, so it is highly attractive to be used in investigations of proteins involved in multimeric protein complexes.