Part:BBa_K2314831
Pmini is a very short constitutive promoter in yeast, which is only 116bp, but with a good strength.
"Most of native yeast promoters can stretch hundreds of base pairs. Specifically, a single-gene circuit carrying a 1.5kb gene requires an additional 1kb of regulatory DNA (between the promoter and terminator) for appropriate expression, thus increasing the DNA cargo load by over 60%. However, this problem hasn’t been focused widely. With the development of synthetic biology, creating short but strong promoter is more important. Pmini is a short but strong constitutive promoter in yeast. It consists three main elements. Core element determines the shortest length required for transcription and serves as a platform for hybrid promoter technology. This core element scaffold was built on distinct, essential sequences for promoter function—a TATA box with consensus sequence of TATAWAWR24 followed by a transcription start site (TSS) with consensus sequence of A(Arich)5NYAWNN(Arich)6. UAS element contains transcription factor-binding sites (TFBS) and can aid in RNAP stabilization to enhanced transcription rates. The AT-rich spacer containing 30 nucleotides has a better performance.[1] We used Pmini promoter with synthetic terminator Tmini to construct a “Mini system“ by constructing four circuits. We wanted to compare the expression of “Mini system” with that of the common “CYC1 system” which contains Pcyc1 and Tcyc1.
[1]Redden H,Alper HS,The development and characterization of synthetic minimal yeast promoters[J],Nature Communication,2015,6 : 7810 "
In short,Pmini is a very short constitutive promoter in yeast, which is only 116bp in length, but with strong expression.
the expression level of mini system:
the RT-PCR result of mini system:
the structure of pmini:
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]
//chassis/eukaryote/yeast
//promoter
chassis | yeast |