Composite

Part:BBa_K3905010

Designed by: Peter Heywood, Harry Weiniger   Group: iGEM21_City_of_London_UK   (2021-09-13)
Revision as of 17:58, 13 September 2021 by Registry (Talk | contribs)

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T7 / gen3 210-3p-517-5p AND-Gate / Luciferase / Double Terminator

This composite part is used for testing the presence of hsa Micro-RNA 517-5p and 210-3p, using AND-gate logic. Both miRNAs must be present to activate this switch. It has a T7 promoter for expression of functional RNA (Part:BBa_K1614000). This is followed by the code for our toehold switch (Part:BBa_K3905006). This toehold switch can act as a riboregulator to inhibit translation until both micro-RNA 210-3p and 517-5p are present. Both of these microRNAs bind to an anti-miRNA (part BBa_K3905007) which forms a complex that presents both target microRNAs as a trigger to the toehold switch. This triggers the toehold switch to unravel, exposing the RBS and start codon, allowing translation of a protein. This protein should be a reporter protein so that the activity of the toehold switch can be detected and presence of the microRNAs can be confirmed. Our chosen reporter protein is luciferase (Part:BBa_I712019). When Luciferin is added, luciferase will catalyse a reaction that produces light. This light can be detected with a luminometer to affirm a positive test.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 44
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 44
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 44
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 108
    Illegal SapI.rc site found at 932


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Categories
Parameters
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