Difference between revisions of "Part:BBa K3110051:Design"

(→‎Design Notes)
(→‎Design Notes)
Line 13: Line 13:
 
<li>A strong RBS: [[Part:BBa_B0034]]
 
<li>A strong RBS: [[Part:BBa_B0034]]
 
<li>Gene Construct: YebF-IL12-FLAG ([[Part:BBa_K3110050]])
 
<li>Gene Construct: YebF-IL12-FLAG ([[Part:BBa_K3110050]])
−
<li>Double terminator: [[Part:BBa_I746916]]
+
<li>Double terminator: [[Part:BBa_B0015]]
 
</ul>
 
</ul>
  

Revision as of 21:18, 21 October 2019


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:

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