Composite

Part:BBa_K2715003:Design

Designed by: Daniel Partridge   Group: iGEM18_Nottingham   (2018-10-05)


Clostridial promoter Pcdi_tcdB with its native 5' UTR and RBS, driving a GFP reporter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 959
    Illegal SapI.rc site found at 139


Design Notes

This composite part is composed of the native clostridial promoter driving expression of the toxin gene tcdB, sub-divided into the promoter region itself BBa_K2715013, the 5’ UTR region BBa_K2715023, and the RBS BBa_K2715024, driving expression of GFP taken from BBa_E0040. An NdeI restriction site was included between the RBS and GFP coding sequence, so that the reporter gene can be easily swapped by performing an NdeI SpeI digest.


Source

The promoter is taken directly from Clostridium difficile upstream of the toxin B gene. The RBS and 5' UTR are also taken from Clostridium difficile upstream of the toxin B gene. GFP is sourced from the jellyfish Aequeora victoria wild-type genome sequence BBa_E0040.