Composite

Part:BBa_K3755014:Design

Designed by: Kaijun Wang   Group: iGEM21_ShanghaiTech_China   (2021-10-01)


NFAT-RE+minP+EGFP


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]


Design Notes

minP has vert low basal activity. So EGFP won't be expressed obviously unless activated NFAT binds to NFAT-RE to enhance the downstream expression.

Source

NFAT reponse element and minimal TATA-box promoter were obtained from pGL4.30 with PCR. EGFP was cloned from pEGFP-vicilin.

References

[1]Weaver JR, Good K, Walters RD, Kugel JF, Goodrich JA. Characterization of the sequence and architectural constraints of the regulatory and core regions of the human interleukin-2 promoter. Mol Immunol. 2007 Apr;44(11):2813-9. doi: 10.1016/j.molimm.2007.01.027. Epub 2007 Mar 6. PMID: 17337059; PMCID: PMC1924494.
[2]Zhang W, Takahara T, Achiha T, Shibata H, Maki M. Cellular Ca2+-Responding Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-palindromic NFAT-Response Elements. Methods Mol Biol. 2019;1929:95-109. doi: 10.1007/978-1-4939-9030-6_7. PMID: 30710269.