Regulatory
Pmini

Part:BBa_K2314831

Designed by: Lei Zhu   Group: iGEM17_OUC-China   (2017-10-22)
Revision as of 15:31, 1 November 2017 by A357034555 (Talk | contribs)

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(BBa_K2314608) to construct a minimal genetic regulatory element “MINI-GRE“ combination. We also selected a commonly used promoter CYC1 and terminator CYC1, and contrusted four circuits to measure the performance of MINI-GRE combination.

[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: https://parts.igem.org/File:Abs600_of_strains_with_different_promoter-terminator_pairs.png#file

the RT-PCR result of mini system:

T--OUC-China--miniRT-PCR.png

the structure of pmini:

T--OUC-China--MINIP.jpg


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]


[edit]
Categories
//awards/basic_part/nominee
//chassis/eukaryote/yeast
//promoter
Parameters
chassisyeast