Part:BBa_K2550001:Design

T7 promoter with a trigger sequence to match the toehold sequence of part BBa_K2550000

Assembly Compatibility:

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

  Illegal NheI site found at 104
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

The RNA sequence was obtained from the Georgia Institue of Technology that was number 2 of the 144 generation orthogonal toehold switch collection from the Collins Paper titled, Toehold Switches: De-Novo-Designed Regulators of Gene Expression.

Design Notes

This part is meant to be used as a proof of concept for the K2550000 Toehold switch. It needs to be ligated into a high copy number plasmid that has a different antibiotic resistance than the Toehold switch plasmid in order to allow the E. coli cell to recognize two distinct plasmids to copy each plasmids at proportional amounts. Lambert iGEM used pSB6A1 for the trigger in our own trials. Dual transformation with both plasmids will result in blue coloration from the LacZ gene.

Source

DNA sequences to build the trigger part was obtained from the Styczynski Lab at Georgia Institute of Technology

References

Alexander A. Green, Pamela A. Silver, James J. Collins, Peng Yin, Toehold Switches: De-Novo-Designed Regulators of Gene Expression, Cell, Volume 159, Issue 4, 2014, Pages 925-939, ISSN 0092-8674, https://doi.org/10.1016/j.cell.2014.10.002