Composite

Part:BBa_K2332040

Designed by: Paola Handal   Group: iGEM17_UCL   (2017-10-20)
Revision as of 13:59, 25 October 2017 by Paola handal (Talk | contribs)


Blue light inducible expression of Intimin'-GFP-SpyTag (Pblind Intimin'-GFP-SpyTag)

This construct displays SpyTag fused to GFP on the cell surface of E. coli cells under the control of Pblind, a blue light inducible promoter. Cells must also express EL222 (BBa_K2332004). Upon blue-light (465nm) exposure, EL222 dimerises and binds to its DNA-binding region within Pblind to recruit RNAP and induce transcription of Intimin'-GFP-SpyTag.

Intimin' is a truncated version of Intimin (EaeA intimin), an outer membrane surface protein, 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 under blue light induction.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 2844
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 2844
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 2844
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 2844
    Illegal NgoMIV site found at 880
    Illegal NgoMIV site found at 1621
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2715


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