Plasmid

Part:BBa_K4212049:Design

Designed by: Yuancheng Ding   Group: iGEM22_Imperial_College_London   (2022-09-30)
Revision as of 11:23, 9 October 2022 by Helen737 (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


Ger1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3591
    Illegal PstI site found at 606
    Illegal PstI site found at 3542
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3591
    Illegal PstI site found at 606
    Illegal PstI site found at 3542
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3591
    Illegal BglII site found at 3123
    Illegal XhoI site found at 713
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3591
    Illegal PstI site found at 606
    Illegal PstI site found at 3542
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3591
    Illegal PstI site found at 606
    Illegal PstI site found at 3542
    Illegal NgoMIV site found at 66
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2778


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. However, we planned to engineer the receptor to make it more sensitive to N-acetylglucosamine (NAG) - the chitin monomer. The previous iGEM team iGEM17_SZU-China proved the superior transcription efficiency of this promoter. tL3S2P21 is a strong terminator, which helps to increase the production of desired protein. We included the operon for germinant receptor GerA because GerA is responsive to L-alanine and responsible for B.subtilis germination. We incorporated GerA into our Golden Gate construct to create the GerAK chimera, as the model our dry lab predicted.


Source

Synthetic construct.

References

[1] https://parts.igem.org/Part:BBa_K2232020 [2] Ramírez-Guadiana, F.H., Meeske, A.J., Wang, X., Rodrigues, C.D.A. & Rudner, D.Z. (2017) The Bacillus subtilis germinant receptor GerA triggers premature germination in response to morphological defects during sporulation. Molecular Microbiology. 105 (5), 689–704. doi:10.1111/mmi.13728.