Composite

Part:BBa_K1800000:Design

Designed by: Laura Lee Irvin   Group: iGEM15_Georgia_State   (2015-08-25)
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Mambalgin-1 for E. coli


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

For the design, we first codon optimized the Mambalgin-1 sequence for Escherichia coli using the IDT codon optimization tool. We added RFC 10 prefix and suffix and also included a start codon in the beginning of the sequence. Also included in the construct was a prokaryotic RBS, an IPTG inducible promoter, and Myc and 6xHis epitopes. This fusion protein was designed specifically for easy expression in E. coli.

Source

Mambalgin-1 sequence (BBa_k1110003) + codon optimization for Escherichia coli, RBS (BBa_j61101), lacI Promoter (BBa_R0011), MYC and 6xHis tag was copied from the pGAPZα vector sequence. This was synthesized by IDT.

References

Diochot, S., Baron, A., Salinas, M., Douguet, D., Scarzello, S., Dabert-Gay, A. S., ... & Lingueglia, E. (2012). Black mamba venom peptides target acid-sensing ion channels to abolish pain. Nature, 490(7421), 552-555.