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. | ||
− | < | + | <img src="https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg>https://static.igem.org/mediawiki/2015/d/dd/MAMBA.E.coli_Western_blot.THISONE.jpg> |
===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 22:57, 26 September 2015
Mambalgin-1 for E. coli
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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=">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.