Composite

Part:BBa_K2332052

Designed by: Paola Handal   Group: iGEM17_UCL   (2017-10-21)
Revision as of 23:38, 22 October 2017 by Paola handal (Talk | contribs)

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Intimin'-GFP-SpyTag (constitutive)

This construct displays GFP fused with SpyTag on the cell surface of E. coli cells constitutively. Intimin' (159kDa) is a truncated version of Intimin (EaeA intimin) which has shown to display proteins of similar size to GFP fused to its N terminal. This construct allows to directly observe through fluorescence microscopy the functionality of Intimin for cell surface display. It can be also used in combination with cells expressing our SpyCatcher construct variants to analyse their co-localization. SpyTag (13 amino acids) and SpyCatcher (138 amino acids, 15 kDa) are protein binding partners that originate from CnaB2 (immunoglobulin-llike collagen adhesin domain) of the FbaB protein, found in the invasive strains of S. pyogenes.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 2826
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 2826
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 2826
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 2826
    Illegal NgoMIV site found at 862
    Illegal NgoMIV site found at 1603
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2697


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Categories
Parameters
None