Composite
Part:BBa_K3110051:Design
Designed by: Amartya Pal Group: iGEM19_IISER_Tirupati (2019-10-16)
lldRO1-J23117-lldRO2 Strong RBS YebF-IL12
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 78
Illegal NheI site found at 101 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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