Part:BBa_K3600000

HSP12 promoter part plasmid

HSP12 promoter part plasmid was made in order to create a part plasmid which contains the HSP12 promoter and is compatible with the Yeast Toolkit. To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter.These linking sequences contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible.Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.

Usage and Biology

The HSP12 gene encodes for a heat-shock protein in Saccharomyces Cerevisiae that is activated by the high-osmolarity glycerol (HOG) pathway.¹ The HSP12 promoter is regulated by the transcription factor MSN2/4.</span> Using the yeast toolkit for modular assembly ² the HSP12 promoter plasmid was used as a part of type II.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 308
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 326
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 308
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 308
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 57
Illegal BsaI.rc site found at 578

Measurements

References

[1] Varela, J. C., Praekelt, U. M., Meacock, P. A., Planta, R. J. & Mager, W. H. The Saccharomyces cerevisiae HSP12 gene is activated by the high-osmolarity glycerol pathway and negatively regulated by protein kinase A. Molecular and Cellular Biology 15, 6232–6245 (1995).

[2] 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).

[3] Martínez-Pastor, M. T. et al. The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). The EMBO Journal 15, 2227–2235 (1996).