Generator

Part:BBa_K1688007:Design

Designed by: Fredrik Lindeberg   Group: iGEM15_Uppsala   (2015-09-15)


ModLac laccase with His-tag and HlyA export tag (inc RBS and J23110 promoter)


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 310
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The modified CueO was synthesized with iGEM prefix and suffix, RBS(BBa_B0034) and a M-scar after the RBS in order to guarantee the function of the RBS. The gene is followed by a KpnI restriction site with glycines and serines on each side, which are small amino acids which increase the flexibility of the protein. This is needed for the export tag HlyA , to make it easier for the resulting signal protein to be folded properly. A second KpnI restriction site is put after the HlyA so that the export tag can be removed easily. It has then two stop codons, and a terminator at the end.


Source

D439A/M510L CueO mutant sequence received from Professor Kunishige Kataoka and synthesized with His-tag and HlyA export tag.

References