Composite

Part:BBa_K3905008

Designed by: Peter Heywood, Harry Weiniger   Group: iGEM21_City_of_London_UK   (2021-09-13)
Revision as of 17:44, 13 September 2021 by Registry (Talk | contribs)

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T7 / gen2 517-5p Toehold Switch / Luciferase / Double Terminator

This composite part is used for testing the presence of hsa Micro-RNA 517-5p. 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_K3905004). This Toehold Switch is activated by hsa-MiRNA 517-5p. The micro RNA 517-5p alone acts as the trigger for the toehold switch and causes the riboregulator to unfold, exposing the RBS and start codon. Placing a reporter protein downstream of this part will mean that when miRNA 517-5p is present in high enough quantities, the protein will be translated and the reporter protein can be observed. 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 41
    Illegal XbaI site found at 79
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 41
    Illegal XbaI site found at 79
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 41
    Illegal XbaI site found at 79
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 922


[edit]
Categories
Parameters
None