Composite

Part:BBa_K2715026:Design

Designed by: Daniel Partridge   Group: iGEM18_Nottingham   (2018-10-08)


Constitutive clostridial promoter Pcsp_fdx (BBa_K2715011), strong RBS and GusA 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
    COMPATIBLE WITH RFC[1000]


Design Notes

In this composite part we've added a strong RBS BBa_K2715020, shown to function in Gram-positive and Gram-negative organisms, downstream of the BBa_K2715011 clostridial promoter, driving expression of GusA taken from BBa_K330002. An NdeI restriction site was included between the RBS and GusA 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 sporogenes upstream of the ferredoxin gene. The RBS and 5’ UTR are also taken from Clostridium sporogenes upstream of the ferredoxin gene. The GusA protein is taken from E. coli.