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Generator

Part:BBa_K1404008:Design

Designed by: Juliette Paillet, Alexandre Duprey, Emiel van der Kouwe   Group: iGEM14_INSA-LYON   (2014-09-24)

p70-CsgA-His*2, double His-tagged curli generator


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

If you intend to use this part, be aware that CsgA might be toxic to E. coli cells in large quantities. We were unable to clone csgA in front of the strong promoter J23119 or the weaker promoter J23114 ,

Source

A strong RBS desgined by team Paris Bettencourt 2009 and the coding region for the csgA gene as found in the genome of the E. coli strain MC4100 were synthesized by Genecust. An illegal restriction site was removed by introducing a silent mutation. A sequence coding for (Gly-Gly-Gly-Gly-His-His-His-His-His-His-Gly-Gly-Gly-Gly-His-His-His-His-His-His) was added at the end of the sequence. This direct synthesis was then cloned downstream of the K342000 promoter by standard assembly.

References