Plasmid

Part:BBa_K3600009

Designed by: yasmine koubaa   Group: iGEM20_EPFL   (2020-10-27)
Revision as of 01:54, 28 October 2020 by Anas (Talk | contribs)


prGSH1-yomScarlet-I-tADH1


This cassette was assembled from 6 part plasmids via Golden Gate assembly using the BsmbBI restriction enzyme. The two connectors (pYTK002 and pYTK072), as well as the terminator (pYTK053) and the Ampicilin resistance/backbone (pYTK095) part plasmids are part of the YTK toolkit developed by Lee et al.1 and can be ordered online. The promoter part plasmid (BBa_K3600003) and the reporter part plasmid (BBa_K3600010) were synthesized by us.

Usage and Biology

This cassette contains the GSH1 promoter upstream of a reporter gene (mScarlet-i). GSH1 is a stress response gene in s.cerevisiae that is regulated by the YAP1 master regulator2. This cassette is then assembled with a part plasmid containing Lys2 homology sequences (BBa_K36000012) in another Golden Gate BsmBI assembly, to give a final integration cassette (BBa_K3600015).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Unknown
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Unknown
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Lee, M. E., DeLoache, W. C., Cervantes, B. & Dueber, J. E. A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synth. Biol. 4, 975–986 (2015).

[2] Gomes DS, Fragoso LC, Riger CJ, Panek AD, Eleutherio EC. Regulation of cadmium uptake by Saccharomyces cerevisiae. Biochimica et Biophysica Acta (BBA)-General Subjects. 2002 Oct 10;1573(1):21-5.

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