Composite

Part:BBa_K1800000:Design

Designed by: Laura Lee Irvin   Group: iGEM15_Georgia_State   (2015-08-25)
Revision as of 07:07, 29 October 2016 by Hbowman11 (Talk | contribs)

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

Mambalgin peptide coding sequence was retrieved from supplementary materials in Diochot, Sylvie, et al. "Black mamba venom peptides target acid-sensing ion channels to abolish pain." Nature 490.7421 (2012): 552-555.