Composite

Part:BBa_K2996501:Design

Designed by: Shifeng Jiang   Group: iGEM19_SJTU-BioX-Shanghai   (2019-10-03)


Terminator put downstream of zinc finger


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 304
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We wanted a relatively strong terminator that can stop the transcription of DBD protein,thus lowering the leakage in the system,so we chose the terminator BBa_B0010 as the terminator.We designed new PCR primers to extend the original DNA sequence of DBD protein(BBa_K2996500),adding the DNA sequence of BBa_B0010 to the sequence.

Source

The subparts of this composite part were BBa_K2996500 and BBa_B0010.BBa_B0010 was added through one additional PCR on the basis of BBa_K2996500.

References

[1]Lohmueller, J. J., Armel, T. Z. & Silver, P. A. A tunable zinc finger-based framework for Boolean logic computation in mammalian cells. Nucleic Acids Research 40, 5180--5187 (2012);

[2]Burz, DS (Burz, DS); Rivera-Pomar, R (Rivera-Pomar, R); Jackle, H (Jackle, H); Hanes, SD (Hanes, SD).Cooperative DNA-binding by Bicoid provides a mechanism for threshold-dependent gene activation in the Drosophila embryo. EMBO JOURNAL 17, 5998-6009(1998)