Coding
ModLac

Part:BBa_K1688020:Design

Designed by: Fredrik Lindeberg   Group: iGEM15_Uppsala   (2015-09-15)
Revision as of 19:17, 15 September 2015 by Registry (Talk | contribs)

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ModLac laccase with His-tag and HlyA export tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 249
  • 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. Two Nsil restriction sites were put before and after a 6xHis-tag so that the enzyme could be purified with an IMAC and so that the tag could be removed. 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