Coding
Part:BBa_K2332010:Design
Designed by: Paola Handal Group: iGEM17_UCL (2017-10-16)
Intimin'-SpyTag
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 801
Illegal NgoMIV site found at 1542 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We decided to fuse SpyTag to the N-terminal of a truncated version of Intimin as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins (Wentzel et al., 2001). We also included a linker between Intimin and SpyTag to facilitate mobility of SpyTag on the cell surface.
Source
Intimin protein sequence obtained from UniProtKB: [http://www.uniprot.org/uniprot/P43261#sequences P43261], reverse translated and codon optimised for E. coli. Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001. SpyTag was obtained from: BBa_K1159201