Difference between revisions of "Part:BBa K3600006"

Revision as of 01:07, 28 October 2020

prHSP12-yomScarlet-I-tADH1

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.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. By integrating this cassette into yeast cells, they will produce a fluorescent signal when they are under stress.

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.

@@ Line 2: / Line 2: @@
 __NOTOC__
 <partinfo>BBa_K3600006 short</partinfo>
 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.
 The promoter part plasmid (BBa_K3600000) and the reporter part plasmid (BBa_K3600010) were synthesized by us.