Regulatory

Part:BBa_K5159005

Designed by: Tianyue WU   Group: iGEM24_HKUST   (2024-09-26)

Yarrowia lipolytica TEF promoter with intron

Yarrowia lipolytica native promoter for Transcriptional Elongation Factor-1α (TEF) is one of the most common strong constitutive promoter used in the chassis, and was previously reported, tested and improved by past teams (see Part:BBa_K2117000).

As it is identified in the genome of Yarrowia lipolytica, a native ribosome-binding site (RBS) is located at the 3’ end of the promoter, sufficient for its proper function, so adding an extra RBS sequence behind is not needed when applying this promoter, which can hinder the proper function of the native RBS sequence (Ouellet et al, 2023).

This part sequence includes a 122 bp native intron which can increase the promoter strength by 17 folds (Stephanopoulos et al, 2013). The mechanism of how this intron increases the promoter strength is still not elucidated. However, the phenomenon of intron improving the native promoter strength is not rare in Yarrowia lipolytica, as around 15% of the native promoters identified include an intron that can affect the promoter strength (Ji et al, 2022). Introns can also be transferred and attached to other identified promoters for tuning the promoters’ strength, which presents one of our future strategies for optimizing inducible promoters in Yarrowia lipolytica.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2

Reference

[1] Benjamin Ouellet, A.M. Abdel-Mawgoud (2023). Strong expression of Cas9 under a new 3′-truncated TEF1α promoter enhances genome editing in Yarrowia lipolytica. Current Research in Biotechnology, Volume 6, 100147. https://doi.org/10.1016/j.crbiot.2023.100147

[2] Mitchell Tai, Gregory Stephanopoulos (2013). Engineering the push and pull of lipid biosynthesis in oleaginous yeast Yarrowia lipolytica for biofuel production. Metabolic Engineering, Volume 15, 1-9. https://doi.org/10.1016/j.ymben.2012.08.007

[3] Mei-Li Sun, Tian-Qiong Shi, Lu Lin, Rodrigo Ledesma-Amaro, Xiao-Jun Ji (2022). Advancing Yarrowia lipolytica as a superior biomanufacturing platform by tuning gene expression using promoter engineering. Bioresource Technology, 347, 126717. https://doi.org/10.1016/j.biortech.2022.126717


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