Composite

Part:BBa_K2974101:Design

Designed by: Janet Standeven, Abby Bell, Ashtyn Cauffman, Monica Cho   Group: iGEM19_Lambert_GA   (2019-10-16)


Medium Promoter (BBa_J23106) Toehold GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 728


Design Notes

Using SnapGene software, the LacZ Reporter Gene (BBa_K2550201) was identified and deleted from the T7 Promoter Toehold Ribosome Switch with LacZ expression (BBa_K2550000), then replaced with and replaced with enhanced Green Fluorescent Protein (BBa_E0040). Additionally, promoter BBa_J23106 was inserted in the T7 Promoter Toehold Ribosome Switch with LacZ expression (BBa_K2550000), replacing the BBa_J23100 promoter. The De-Novo Toehold Switch sequence (BBa_K2550100) was obtained from the Collins Lab at the Massachusetts Institute of Technology and was visualized through the NuPACK software.

T--Lambert GA--Snapgene2.png

Figure 1: Designed BBa_J23106 Toehold eGFP construct in the SnapGene software.



T--Lambert GA--NuPACK1.png

Figure 2: Visualized DeNovo toehold switch using the NuPack software, originally designed in the Collins lab and further obtained from the Styczynski Lab at the Georgia Institute of Technology.


Source

Green, A., Silver, P., Collins, J., & Yin, P. (2014). Toehold Switches: De-Novo-Designed

References