Composite

Part:BBa_K2542012

Designed by: Yaoqiang Zheng   Group: iGEM18_NEFU_China   (2018-10-05)


pCYC-Stemloop1-EGFP-tCYC1

This is a composite part, and the stem loop which at the downstream of the promoter locks the transcription of EGFP. The stem loop makes EGFP impossible to express. Only when a specific small RNA(BBa_K2542003) binds to the stem loop，which can alter the secondary structure of the stem loop, leading EGFP transcription to be initiated and EGFP to be expressed.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Experiment

We constructed plasmids pesc-ura-stem loop-EGFP-URA & pesc-ura-EGFP-URA，and transform these plasmid into yeast.Then we used a fluorescence microscope to observe the transformed yeast. The results indicate that the stem loop can effectively prevent the expression of downstream genes.

(A) Photograph of yeasts without stem loop in the light field. (B) Photograph of yeasts without stem loop in the dark field. (C) Photograph of yeasts with stem loop in the light field.(D) Photograph of yeasts with stem loop in the dark field.


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