Composite

Part:BBa_K3725040

Designed by: Andrew Bae, Pranav Kanthala, Monica Cho, Janet Standeven, Megan McSweeney   Group: iGEM21_Lambert_GA   (2021-09-28)
Revision as of 17:06, 20 October 2021 by Michellejing (Talk | contribs)


T7 Fusarium Trigger

Overview

The T7 Phytophthora Trigger is designed to be used in conjunction with the T7 promoter to induce GFP expression for Lambert iGEM’s P. Cryptogea toehold biosensor. When the trigger RNA sequence is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop allowing the reporter protein (GFP) to be expressed, producing green fluorescence. The sequence was designed via NUPACK using an input code provided by Takahashi et. al. We ordered the insert in a pUCIDT Kan plasmid obtained from their website.

Description Toehold biosensors, which are composed of a switch and trigger, are highly orthogonal riboregulators that activate translation in response to a specific RNA sequence. The switch is composed of a hairpin loop structure that represses translation through its complementary bases in between the ribosomal binding site and the start codon, which is followed by a linker sequence. Once the toehold is exposed to the trigger sequence, the complementary base pairs on the trigger will bind to the toehold, which exposes the ribosomal binding site. RNA polymerase can then bind to the RBS and initiate translation of the reporter protein.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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