Composite

Part:BBa_K4779002

Designed by: Ziqi Mi   Group: iGEM23_Nanjing-BioXstem   (2023-09-26)
Revision as of 19:15, 11 October 2023 by LinXMz (Talk | contribs)


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

It is a copper ion reaction reporting pathway in Saccharomyces cerevisiae. We employed a positive feedback regulation with Ste5ΔN-CTM for signal amplification. In addition, we utilized the prm1 promoter for both GFP and Ste5ΔN-CTM expression.

Construction

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.


nanjing-bioxstem-cmps.png

Characterization

We subjected the engineered yeast strain BY4741-pRS415-CMPS samples to a copper ion gradient treatment and quantitatively measured their fluorescence intensity at different copper ion concentrations, as depicted in the graph. Within 500 uM, the signal output intensity of the CMPS sensor increased with an ascending copper ion concentration and nearly reached its peak at 50 uM. In comparison to CMPG, which reached its signal output peak at 200 uM, CMPS achieved comparable signal output intensity at lower copper ion concentrations, indicating an improved sensitivity.


nanjing-bioxstem-pprm1-pro2.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    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
    Illegal PstI site found at 49
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2657
    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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2657
    Illegal BglII site found at 55
    Illegal BglII site found at 3105
    Illegal BglII site found at 4413
    Illegal BamHI site found at 510
    Illegal XhoI site found at 8
    Illegal XhoI site found at 4350
  • 23
    INCOMPATIBLE WITH RFC[23]
    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
    Illegal PstI site found at 49
  • 25
    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
    Illegal PstI site found at 49
    Illegal NgoMIV site found at 36
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2213
    Illegal SapI site found at 529


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