Regulatory

Part:BBa_K4140012

Designed by: Ahmed Gamal Mohamed Mattar   Group: iGEM22_AFCM-Egypt   (2022-09-24)


Human U6 promoter


Part Description

A proximal sequence element, which attracts the multisubunit component SNAPc, and a TATA box, which recruits the TATA box-binding protein, TBP, make up the human U6 small nuclear RNA core promoter. Two clearly identified factors are necessary for transcription from the human U6 promoter in addition to SNAPc and TBP. The first is the human homologue of the yeast TFIIIB subunit B" (hB), which is generally necessary for the transcription of RNA polymerase III genes, and the second is the human homologue of the TFIIIB subunit BRF (hBRFU), which is one of two human homologues and is specifically necessary for the transcription of U6-type RNA polymerase III promoters.

Usage

Promoter of the core of the human U6 small nuclear RNA. constructed from For transcribing, two distinctly outlined elements are required. We use a proximal sequence element, which draws the multisubunit component SNAPc, and a TATA box, which draws the TATA box-binding protein, TBP, to control the expression of our regulatory system, that is composed of two domains: the Cas12g protein and gRNA, which is in charge of fine-tuning of our circuit to prevent excessive PAH expression in the therapeutic circuit and beta-galactosidase in the diagnostic circuit as shown in figure 1.

Figure(1) Shows an SBOL demonstrating the usage of human 6U promoter in the regulatory circuit in our whole cel-based biosensor








Experimental Characterization

Capture7.png






This figure shows an experimental characterization of this part as it's validated through gel electrophoresis as it is in lane 4. The running part (ordered from IDT) included Human u6 Promoter - -Kinkturn - CMV Promoter - Cas12g.















References

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]


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Categories
Parameters
None