Composite
CacthlGFP

Part:BBa_K2715001:Design

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


Constitutive clostridial promoter Pcac_thl (BBa_K2715010), strong RBS and 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 797


Design Notes

In this composite part we've added a strong RBS BBa_K2715009, shown to function in Gram-positive and Gram-negative organisms, downstream of the BBa_K2715010 clostridial promoter, driving expression of GFP taken from BBa_E0040. This composite part also includes the 5’ UTR region of the thiolase promoter BBa_K2715019. An NdeI restriction site was included between the RBS and GFP gene, so that the reporter gene can be easily swapped by performing an NdeI SpeI digest.


Source

The promoter is taken directly from Clostridium acetobutylicum upstream of the thiolase gene. The RBS is also taken from Clostridium acetobutylicum upstream of the thiolase gene. GFP is sourced from the jellyfish Aequeora victoria wild-type genome sequence BBa_E0040.