Composite

Part:BBa_K1800000:Design

Designed by: Laura Lee Irvin   Group: iGEM15_Georgia_State   (2015-08-25)
Revision as of 22:57, 26 September 2015 by Jsiemer (Talk | contribs) (→‎Source)

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. <img src="MAMBA.E.coli_Western_blot.THISONE.jpg>https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg>

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.