Difference between revisions of "Part:BBa K3600006"
 
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<partinfo>BBa_K3600006 short</partinfo>
 
<partinfo>BBa_K3600006 short</partinfo>
  
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This cassette was assembled from 6 part plasmids via BsmbI assembly. 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.<html><a href="#Lee2015"><sup>1</sup></a></html> and can be ordered online.
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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.<html><a href="#Lee2015"><sup>1</sup></a></html> and can be ordered online.
 
The promoter part plasmid (BBa_K3600000) and the reporter part plasmid (BBa_K3600010) were synthesized by us.  
 
The promoter part plasmid (BBa_K3600000) and the reporter part plasmid (BBa_K3600010) were synthesized by us.  
  
 
===Usage and Biology===
 
===Usage and Biology===
  
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This cassette contains the HSP12 promoter upstream of a reporter gene (mScarlet-i). HSP12 is a stress response gene in S.cerevisiae that is regulated by the MSN2/4 master regulator<html><a href="#Sadeh2011"><sup>2</sup></a></html>. By integrating this cassette into yeast cells, they will produce a fluorescent signal when they are under stress.
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This cassette contains the HSP12 promoter upstream of a reporter gene (mScarlet-i). HSP12 is a stress response gene in s.cerevisiae that is regulated by the MSN2/4 master regulator<html><a href="#Sadeh2011"><sup>2</sup></a></html>.  
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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_K3600013).
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 01:51, 28 October 2020


prHSP12-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_K3600000) and the reporter part plasmid (BBa_K3600010) were synthesized by us.

Usage and Biology

This cassette contains the HSP12 promoter upstream of a reporter gene (mScarlet-i). HSP12 is a stress response gene in s.cerevisiae that is regulated by the MSN2/4 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_K3600013).

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] Sadeh A, Movshovich N, Volokh M, Gheber L, Aharoni A. Fine-tuning of the Msn2/4–mediated yeast stress responses as revealed by systematic deletion of Msn2/4 partners. Molecular biology of the cell. 2011 Sep 1;22(17):3127-38.