Plasmid

Part:BBa_K3600011

Designed by: yasmine koubaa   Group: iGEM20_EPFL   (2020-10-27)
Revision as of 01:31, 28 October 2020 by Anas (Talk | contribs)


prGLR1-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_K3600004) and the reporter part plasmid (BBa_K3600010) were synthesized by us.

Usage and Biology

This cassette contains the GLR1 promoter upstream of a reporter gene (mScarlet-i). GLR1 is a stress response gene in s.cerevisiae that is regulated by the YAP1 master regulator2.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal XbaI site found at 1193
    Illegal XbaI site found at 1618
    Illegal XbaI site found at 2091
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
    Illegal NotI site found at 1172
    Illegal NotI site found at 3078
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal suffix found in sequence at 2661
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal BglII site found at 1636
    Illegal BglII site found at 2532
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal XbaI site found at 1193
    Illegal XbaI site found at 1618
    Illegal XbaI site found at 2091
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal XbaI site found at 1193
    Illegal XbaI site found at 1618
    Illegal XbaI site found at 2091
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
    Illegal NgoMIV site found at 3016
  • 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] Grant CM, MacIver FH, Dawes IW. Stationary‐phase induction of GLR1 expression is mediated by the yAP‐1 transcriptional regulatory protein in the yeast Saccharomyces cerevisiae. Molecular microbiology. 1996 Nov;22(4):739-46.

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