Part:BBa_K2332014:Design
Blue light inducible expression of Intimin'-SpyCatcher (Pblind Intimin'-SpyCatcher)
- 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 898
Illegal NgoMIV site found at 1639 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We decided to fuse SpyCatcher 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 SpyCatcher to facilitate mobility of SpyCatcher on the cell surface and a HisTag for protein purification. Blue light inducible promoter was designed by Jayaraman P. et al. (2016)
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. SpyCatcher was obtained from: BBa_K1159200. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)