Part:BBa_K3905012
T7 / gen3 210-3p-517-5p AND-Gate / Luciferase / Double Terminator / T7 / gen3 210-3p-517-5p AND-Gate
This composite part codes for the toehold switch and the anti-mirna to test for the presence of hsa micro-RNA 210-3p and 517-5p. 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. The composite part has the same T7 promoter twice, for expression of functional RNA (Part:BBa_K1614000). This is a suitable promoter for this part due to the functional RNAs that are being coded for. There are two different terminators present, the double terminator (Part:BBa_B0015) follows the toehold switch and the standard terminator follows the anti-MiRNA (Part:BBa_B1002).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 108
Illegal SapI.rc site found at 932
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