Composite

Part:BBa_K2332014:Design

Designed by: Paola Handal   Group: iGEM17_UCL   (2017-10-16)
Revision as of 00:02, 23 October 2017 by Paola handal (Talk | contribs) (→‎Design Notes)


Blue light inducible expression of Intimin'-SpyCatcher (Pblind Intimin'-SpyCatcher)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 898
    Illegal NgoMIV site found at 1639
  • 1000
    COMPATIBLE 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

All parts were obtained as synthetic gBlocks Intimin protein sequence obtained from: http://www.uniprot.org/uniprot/P43261#sequences And reverse translated using: http://www.bioinformatics.org/sms2/rev_trans.html Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001 SpyCatcher: BBa_K1159200 Pblind:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5001607/

References