Difference between revisions of "Part:BBa K1800000:Design"
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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.
 
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.
 
This was synthesized by IDT.
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<href="https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg</ref>
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<ref="https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg</ref>
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===References===
 
===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.
 
  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.

Revision as of 22:53, 26 September 2015

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. <ref="MAMBA.E.coli_Western_blot.THISONE.jpg</ref>

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.