Regulatory

Part:BBa_K3054004

Designed by: Dharmik Hitesh Patel   Group: iGEM19_Chalmers-Gothenburg   (2019-10-20)


pTEF1/pCIT2 dual promoter (yeast)


Dual promoter design

A dual promoter system generated by our team that consists of two strong promoters used in our lab from Saccharomyces cerevisiae. These promoters are placed in such a way that they can transcribe two protein sequences in opposite directions. Linker between these two promoters, 30bp of otherwise non-functional DNA, is not complementary to Saccharomyces cerevisiae genome. This helps in primer designing when extracting these promoters from Yeast genome and helps link 2 yeast promoters by fusion PCR. Diagram shows the design of dual promoter in general



Reference: Yasaman Dabirian, Xiaowei Li, Yun Chen, Florian David, Jens Nielsen, and Verena Siewers. Expanding the Dynamic Range of a Transcription Factor-Based Biosensor in Saccharomyces cerevisiae. ACS Synthetic Biology 2019 8 (9), 1968-1975. DOI: 10.1021/acssynbio.9b00144




Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1427
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1427
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1427
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1427
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 241


[edit]
Categories
Parameters
None