Composite

Part:BBa_K2557029

Designed by: Wang Chunfeng   Group: iGEM18_NAU-CHINA   (2018-10-09)


TetO-EF1α promoter -EGFP

The EF1α promoter and EGFP coding sequences were ligated in sequence to characterize promoter strength.

Sequence and Features

Usage and Biology

This composite part can transduce the input signal of the upstream TEV protease into the fluorescent signal of Enhanced GFP. Here, we characterized the combination of the TetO-EF1α promoter.

Characterization

Fig.1 Inhibition of tetR on promoter with tetO sequence (A)Fluorescence microscope observation of HEK 293T only transfect with plasmids containing promoters with tetO sequence. (B)Fluorescence microscope observation of HEK 293T transfect with plasmids containing promoters with tetO sequence and tetR.

Fig. 1A shows that the strength of the three promoters in HEK 293T cells is from strong to weak: Ubc > EF1 α > miniCMV. Fig. 1B shows that TetR has obvious inhibitory effects on the three promoters downstream of TetO.

References

1. Ramos, J. L. et al. The TetR Family of Transcriptional Repressors The TetR Family of Transcriptional Repressors. Microbiol. Mol. Biol. Rev. 69, 326–356 (2005).



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 499
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 505
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/eukaryote
Parameters
None