Part:BBa_K5121019

rebA C-teriminal LPETG

Biology

RebA encodes an 18 kDa protein which forms one of the main structural components of type 51 R bodies. This part is based on the wildtype rebA part — BBa_K2912000 — uploaded by SZU-China in 2019, however has been modified to be compatible with sortase A-mediated conjugation. Sortase A is a bacterial transpeptidase enzyme that ligates C-terminal LPXTG motifs with N-terminal polyglycine motifs (Theile et al., 2013).

Our group added an LPETG peptide sequence to the C-terminus of the rebA monomer. This modification facilitates conjugation of proteins/small molecules containing an N-terminal polyglycine motif to the C-terminus of rebA in the presence of sortase A. Using this approach, we conjugated the fluorescent protein mNeonGreen with an N-terminal polyglycine sequence (BBa_K5121012) to R-bodies containing the modified rebA monomer. See the design page for more information.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 803
  Illegal NheI site found at 865
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 745
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

Theile, C. S., Witte, M. D., Blom, A. E. M., Kundrat, L., Ploegh, H. L., & Guimaraes, C. P. (2013). Site-specific N-terminal labeling of proteins using sortase-mediated reactions. Nature Protocols, 8(9), 1800–1807.