Part:BBa_K4779000
CMPG:A yeast copper-induced reporter pathway with prm1 promoter
This is a copper ion-responsive signaling pathway in Saccharomyces cerevisiae based on MAPK system. When the cells were induced by copper ions, the circuit with prm1 promoter activates GFP expression to indicate the copper ion concentration.
Construction
We obtained the gene sequences of the basic parts from IGEM, such as the promoter of Cup1 (BBa_K945002), MFα2 (BBa_K110005), the promoter of prm1 (BBa_K1346004), green fluorescent protein (GFP) (BBa_K3112009), and the terminator CYC1 (BBa_K4278703), and sent them to GENEWIZ (Suzhou Genewiz Biotechnology Co., Ltd.) for synthesis. The new composite part (CMPG) (BBa_K4779000) is also synthesized.
Characterization
We treated the engineered yeast strain BY4741-pRS415-CMPG samples with a copper ion gradient, and detected them using a flow cytometer. The quantitative measurements of fluorescence intensity at various copper ion concentrations are shown in the graph. Within 200 uM, the sensor's signal output strength increases with an increase in copper ion concentration.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 279
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Illegal PstI site found at 49 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 343
Illegal SpeI site found at 523
Illegal PstI site found at 49 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 55
Illegal BamHI site found at 510
Illegal XhoI site found at 8 - 23INCOMPATIBLE WITH RFC[23]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 PstI site found at 49 - 25INCOMPATIBLE WITH RFC[25]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 PstI site found at 49
Illegal NgoMIV site found at 36 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2213
Illegal SapI site found at 529
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