Plasmid

Part:BBa_K4212045:Design

Designed by: Yuancheng Ding   Group: iGEM22_Imperial_College_London   (2022-09-30)


SDP2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 150
    Illegal PstI site found at 910
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 91
    Illegal PstI site found at 910
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 150
    Illegal PstI site found at 910
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 150
    Illegal PstI site found at 910
    Illegal NgoMIV site found at 66
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PsspB is a promoter, which can aid the expression of gerA. GerA is one of the germinant receptors of B.subtilis, and it can respond to the presence of L-alanine and increase the germinantion rate of B.subtilis. The previous iGEM team iGEM17_SZU-China proved the superior transcription efficiency of this promoter. The D15 gene encodes an nuclease, which helps to produce nicks at DNA (Moyer & Rothe, 1977). We included it in our self-digesting plasmid construct. Spacers were created to maximize the ligation fidelity in the final assembly.

Source

Synthetic construct.

References

[1] https://parts.igem.org/Part:BBa_K2232020 [2] Moyer, R.W. & Rothe, C.T. (1977) Role of the T5 gene D15 nuclease in the generation of nicked bacteriophage T5 DNA. Journal of Virology. 24 (1), 177–193. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC515921/. [3] https://goldengate.neb.com/#!/help