Composite

Part:BBa_K1354000:Design

Designed by: Devora Najjar, Shoshana Sigal   Group: iGEM14_Cooper_Union   (2014-10-03)


UV Promoter with Phage Activator


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

The sequence we used included the four parts listed above. We also added in extra enzymes sites so that the UV promoter could be switched out easily for any other promoter. Before the UV promoter there is an XhoI site and after it there is a KpnI site. The Delta Activator from Phage phiR73 was chosen based on Cambridge's 2009 iGEM project. In that, they measured this phage activator to be the strongest of the ones they worked with. In order to avoid any issues with our two plasmid system, we chose the strongest one to make sure the connection between the two plasmids would work. We chose to work with a UV promoter because of the ease to work with it. All that was necessary to turn the promoter on was a five second UV shock. One limitation in this is that the UV light can also kill many of the cell colonies; a UV shocked plate would have fewer colonies than a control plate that wasn't shocked.

Source

References