Device
Part:BBa_K2008003:Design
Designed by: Rachelle Varga Group: iGEM16_UofC_Calgary (2016-10-08)
pVeg->sec-TD1-KSCI-BBI-GFP
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 925
Design Notes
Bacillus subtilis-specific components were used (strong RBS and constitutive promoter) to produce high levels of expression of the BBI peptide fused to GFP for visualization. A secretory signal peptide sequence was included to allow for secretion of the peptide directly into surrounding media. A transdermal tag allowed for diffusion of the peptide across the skin. Codons were optimized for B. subtilis. Additional amino acids were included to increase solubility of the desired peptide product.
Source
Members of the lentil family produce the full BBI protein, and the genetic sequence for the peptide was sourced originally from genomic DNA.