Regulatory

Part:BBa_K3423001

Designed by: Linsheng Zhang   Group: iGEM20_SUSTech_Shenzhen   (2020-10-17)


Anti-CRISPR (acr) promoter region

This Anti-CRISPR promoter is a constitutive promoter that can be inhibited by protein Anti-CRISPR-associated 1 (Stanley et al., 2019).

Protein anti-CRISPR-associated 1 (Aca1) is expressed by T7 promoter, Aca1 will subsequently inhibit the anti-CRISPR promoter, so the gene expression downstream anti-CRISPR will be inhibited until Navitoclax (a compound that can inhibit Aca1 found by SUSTech_Shenzhen) is added artificially. Without the inhibition from Aca1, the expression of genes downstream anti-CRISPR promoter will increase significantly. Therefore, protein expression can be regulated artificially.

We characterised the efficiency of this part of promoting a reporter gene in a protein regulation system, the reporter was sufficiently expressed for measurement, acr promoter successfully worked in our composite part BBa_K3423005. Green fluorescent protein is the reporter, the green fluorescence intensity is measured: SUSTech_ShenzhenGFP.png

Reference

Stanley, S. Y., Borges, A. L., Chen, K.-H., Swaney, D. L., Krogan, N. J., Bondy-Denomy, J., & Davidson, A. R. (2019). Anti-CRISPR-Associated Proteins Are Crucial Repressors of Anti-CRISPR Transcription. Cell, 178(6), 1452-1464.e1413. doi:10.1016/j.cell.2019.07.046


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