Part:BBa_K3110051:Design

lldRO1-J23117-lldRO2 Strong RBS YebF-IL12

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 78
  Illegal NheI site found at 101
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

IL12 and YebF was two different parts used by two different iGEM teams. In order to improve the part used by Brazil (BBa_K554005), we have codon optimized it to make it synthesize compatible and attached a Secretory peptide YebF so that IL12 also get secreted along with YebF

So, the whole part contains:

• The Promoter and Operator Region: lldRO1-J23117-lldRO2 (Part:BBa_K1847008)
• A strong RBS: Part:BBa_B0034
• Gene Construct: YebF-IL12-FLAG (Part:BBa_K3110050)
• Double terminator: Part:BBa_B0015

Source

YebF-IL12-FLAG tag was synthesized by Twist Bioscience. lldRO1-J23117-lldRO2 + B0034 and B0015 were obtained as gBlocks from IDT and these 3 parts were joined together using SOEing (Spliced Overlap Extension) PCR.

References

• Zhang, G., Brokx, S., & Weiner, J. H. (2005). Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli. Nature Biotechnology, 24(1), 100–104. doi:10.1038/nbt1174
• Team ETH Zurich 2015