Composite

Part:BBa_K2715114:Design

Designed by: Christopher Humphreys, Daniel Partridge, Lucy Allen, Nemira Zilinskaite   Group: iGEM18_Nottingham   (2018-09-21)


Constitutive E.coli promoter BBa_J23114, strong RBS and GFP reporter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 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 706


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 previously used constitutive E. coli promoter BBa_J23114, driving expression of GFP. 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 J23114 is a derivative of J23119, present in pSB1A2 and sourced from the iGEM registry, the RBS is taken from Clostridium acetobutylicum upstream of the thiolase gene, and GFP is sourced from the jellyfish Aequeora victoria wild-type genome sequence BBa_E0040.