Part:BBa_K1734006

6xHis-T2A-Nanoluc

This part encodes a sequence capable to add a purification tag (6X His) at the 3’ terminus of a desired protein. Furthermore, by using the reporter gene (nanoluc) it’s possible to quantify indirectly the protein concentration. The T2A sequence cleaves the protein [1], producing the protein of interest tagged with HisTag and nanoluc separately This part has been codon optimized for Spodoptera frugiperda (Sf9)

This part has been confirmed by restriction analysis of the pSB1C3 vector

Reference:

[1] Kim JH, Lee S-R, Li L-H, Park H-J, Park J-H, Lee KY, et al. (2011) High Cleavage Efficiency of a 2A Peptide Derived from Porcine Teschovirus-1 in Human Cell Lines, Zebrafish and Mice. PLoS ONE 6(4): e18556. doi:10.1371/journal.pone.0018556

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 29
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 532