Composite

Part:BBa_K3453163

Designed by: Maeva Cherriere   Group: iGEM20_Evry_Paris-Saclay   (2020-10-04)


Rosewood DmTrnL-UAA Trigger 1.3 transcription unit

This part is a transcription unit of the Rosewood DmTrnL-UAA Trigger 1.3 (BBa_K3453063) for sequence-based detection of rosewood. The transcription is controlled by the T7 promoter (BBa_K2150031) and the strong SBa_000587 synthetic terminator (BBa_K3453000).

Usage and Biology

A toehold switch is an RNA–based device containing a ribosome binding site (RBS) and an ATG start codon embedded in a hairpin structure that blocks translation initiation [1]. The hairpin can be unfolded upon binding of a trigger RNA thereby exposing the RBS and the ATG start codon permitting translation of the reporter protein (Figure 1).

T--Evry Paris-Saclay--toehold-switch.png

Figure 1. Toehold switches principle.

This part is a transcription unit of BBa_K3453063 which is a trigger for the Rosewood DmTrnL-UAA Toehold Switch 1.3 (BBa_K3453053) and also contains the switch binding site for the Rosewood DmTrnL-UAA Toehold Switches 2.1 (BBa_K3453054) and 2.2 (BBa_K3453055).

This part proved to be functional: the transcribed rosewood RNA was able to bind to the Rosewood DmTrnL-UAA Toehold Switches 1.3 and 2.2 and a readable output was generated. However, the Rosewood DmTrnL-UAA Toehold Switch 2.1 turned out to be not functional and the interaction with this switch could not be assessed.

Full results are available on the BBa_K3453153, BBa_K3453154 and BBa_K3453155 pages in the registry.

References

[1] Green AA, Silver PA, Collins JJ, Yin P. Toehold switches: de-novo-designed regulators of gene expression. Cell (2014) 159, 925-939.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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