Part:BBa_K3956003
cpTEF_6-I core promoter (synthetic)
This is a constitutive synthetic yeast promoter. In 2019, Decoene et. al constructed short, synthetic core promoters to modulate transcription with cpTEF_6-I being the strongest promoter with an 8-fold protein expression range. The promoter was first used in CCA_San_Diego’s 2021 CollaGene project to initiate the transcription of a bacterial collagen protein in yeast.
Usage and Biology
This sequence should be used right after an upstream activating sequence and a 30 bp spacer sequence (BBa_K3956001). The promoter was designed by Decoene et al., 2019, and includes a transcription start site (TATA-like sequence with base pairs gatttaaa) before a 5’UTR and the open reading frame of our construct.
In our project, as we were expressing a bacterial protein from S. cerevesiae, having a strong promoter was crucial to inducing noticeable expression of our desired scl2 construct. Natural promoters with a high variance across different experiments have a lower predictability and may make their heterologous pathways unfavorable (Decoene et. al, 2019). In other words, synthetic promoters may be optimal for genetic engineering purposes compared to natural promoters, as it reduces the risk of off-target binding when targeting the expression of recombinant constructs. After searching the literature on the topic, we found that the synthetic TEF1-derived promoters documented by Decoene et. al, 2019, fulfilled all of our criteria. As TEF1 is a well-characterized strong promoter in S. cerevisiae, the study authors randomized their elucidated sequence of the TEF1 minimal core promoter. They also discovered that an upstream activating sequence (UAS) around 100 bp in length dramatically improved fluorescence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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