Composite

Part:BBa_K4779002:Design

Designed by: Ziqi Mi   Group: iGEM23_Nanjing-BioXstem   (2023-09-26)


CMPS:A yeast copper-induced reporter pathway with Positive feedback loop in with prm1 promoter


Assembly Compatibility:
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    INCOMPATIBLE WITH RFC[10]
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    INCOMPATIBLE WITH RFC[12]
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    INCOMPATIBLE WITH RFC[21]
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    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2657
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    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal SpeI site found at 3912
    Illegal SpeI site found at 4458
    Illegal PstI site found at 49
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    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2657
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal SpeI site found at 3912
    Illegal SpeI site found at 4458
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    Illegal NgoMIV site found at 36
  • 1000
    INCOMPATIBLE WITH RFC[1000]
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Design Notes

When meticulously designing sequences, one of the critical considerations is how to enhance the sensitivity and efficiency of converting copper ion signals into pheromone signals. According to the literature, Ste5ΔN-CTM has been reported to anchor Ste5 constitutively to the plasma membrane, thereby maintaining the activation of the MAPK pathway. Therefore, in this round DBTL, we utilized Ste5ΔN-CTM to construct a positive feedback loop, where the pheromone-inducible promoter pprm1 controls the expression of Ste5ΔN-CTM protein. This activation enhances the amplification effect of the MAPK pathway, further elevating the sensitivity in detecting pheromone which was produced by pCUP1.


Source

CMPS is a new composite part (BBa_K4779002). We obtained the gene sequences of the promoter of Cup1 (BBa_K945002), MFα2 (BBa_K110005), the promoter of prm1 (BBa_K1346004), green fluorescent protein (GFP) (BBa_K3112009), Ste5ΔN-CTM (BBa_K3384315) and the terminator CYC1 (BBa_K4278703) from IGEM, and sent them to GENEWIZ (Suzhou Genewiz Biotechnology Co., Ltd.) for synthesis.

References