Composite

Part:BBa_K2332025:Design

Designed by: Paola Handal   Group: iGEM17_UCL   (2017-10-20)
Revision as of 00:10, 23 October 2017 by Paola handal (Talk | contribs) (→‎Design Notes)


Intimin'-mutSpyCatcher (Lys31X, for photocaging)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 880
    Illegal NgoMIV site found at 1621
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This construct was designed to not contain any other amber stop codon but at position 31 (Lys31X) (Amberless E. coli must be used to see the effects of photocageing spyCatcher). 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.

Source

SpyCatcher: BBa_K1159200


References