Composite

Part:BBa_K4653907:Design

Designed by: Tangkun Zhu   Group: iGEM23_SZU-China   (2023-10-10)


T7 promoter (lacⅠ repressible) - shRNA (Pme1)-2 - T7 Terminator


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 107


Design Notes

We modified nuclease-deficient Escherichia coli to produce shRNA, and used T7 promoter and lac operon to regulate our expression of shRNA.


Source

After confirming the selection of targets Bcpme1, we searched the cDNA library of B. cinerea according to the sequences or primes provided in the literature, and found the homologous cDNA sequence of B. cinerea. Then, the sequence was input into the National Center for Biotechnology Information (NCBI) website for analysis and prediction, and the CDS sequence of the target gene was input into the total nucleic acid database BLAST to query the homologous similarity of neighboring species. siRNA sequences were designed in non-conserved regions to ensure species-specific and biosafety of our shRNAs.

References