Difference between revisions of "Part:BBa K3453163"

Latest revision as of 21:53, 26 October 2020

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).

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]

@@ Line 6: / Line 6: @@
 ===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 therefore exposing the RBS and the ATG start codon permitting translation of the reporter protein (Figure 1).
+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).
 [[File: T--Evry_Paris-Saclay--toehold-switch.png|600px]]
@@ Line 14: / Line 14: @@
 This part is a transcription unit of [[Part:BBa_K3453063|BBa_K3453063]] which is a trigger for the Rosewood DmTrnL-UAA Toehold Switch 1.3 ([[Part:BBa_K3453053|BBa_K3453053]]) and also contains the switch binding site for the Rosewood DmTrnL-UAA Toehold Switches 2.1 ([[Part:BBa_K3453054|BBa_K3453054]]) and 2.2 ([[Part:BBa_K3453055|BBa_K3453055]]).
-This part proved to be functional: the transcribed rosewood RNA was able to bind to the corresponding toehold switches and a readable output was generated.
+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 [[Part:BBa_K3453153|BBa_K3453153]], [[Part:BBa_K3453154|BBa_K3453154]] and [[Part:BBa_K3453155|BBa_K3453155]] pages in the registry.