Coding

Part:BBa_K511206:Design

Designed by: Grant Robinson   Group: iGEM11_MIT   (2011-09-27)

LacI-miRFF4 Repressor MammoBlock


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1332
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1332
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1332
    Illegal XhoI site found at 1229
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1332
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1332
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Because the FF4 miRNA was designed to bind mRNA in a three-dimensional manner, the FF4 sequence has significant secondary structure. This makes it difficult to design primers that will capture the miRFF4 region. To work around this problem, we suggest using restriction cloning rather than PCR-based methods to isolate either the LacI-miRFF4 part or the miRFF4 region specifically.


Source

This part is a concatenation of a mammalian codon-optimized version of the LacI repressor originally derived from Escherichia coli and the synthetic FF4 miRNA sequence. Sincere thanks go to the Weiss lab for contributing this entry vector to the MIT iGEM 2011 team.

References