Generator

Part:BBa_K638401:Design

Designed by: Matthew Jones   Group: iGEM11_Cambridge   (2011-09-19)


Arabinose inducible TorA Export Tagged Reflectin A1 Poly His generator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 1369
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

We used a TorA sequence from the literature that had been shown to cause GFP to be exported to the periplasm of e.coli. This sequence has also been checked for illegal restriction sites - using the tool A plasmid Editor (ApE) a restriction digest prediction revealed no sites for EcoRI, SpeI, XbaI or PstI.


Source

Parts from the registry, Reflectin A1 DNA from Wendy Crookes, TorA sequence from paper: Export of active green fluorescent protein to the periplasm by the twin-arginine translocase (Tat) pathway in Escherichia coli Joanna D. Thomas1, Richard A. Daniel2, Jeff Errington2, Colin Robinson1,*

The TorA sequence was incorporated into the construct using Gibson Assembly.

References