Difference between revisions of "Part:BBa K3927005"
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===Description=== | ===Description=== | ||
PGK1-tetO is a PGK1 S.cerevisiae promoter with a tet-operon inserted 20 basepairs downstream of it’s TATA box(Figure 1). TetR has been shown to bind to and repress tetO containing synthetic promoter in S.cerevisiae[1]. TetO was placed downstrem of the TATA box in order to construct a promoter that was constitutive unless TetO was expressed in the same cell | PGK1-tetO is a PGK1 S.cerevisiae promoter with a tet-operon inserted 20 basepairs downstream of it’s TATA box(Figure 1). TetR has been shown to bind to and repress tetO containing synthetic promoter in S.cerevisiae[1]. TetO was placed downstrem of the TATA box in order to construct a promoter that was constitutive unless TetO was expressed in the same cell | ||
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===Design=== | ===Design=== | ||
Revision as of 14:01, 18 October 2021
PGK1-TetO
Native PGK1p promoter with TetO operator downstream of TATA box such that promoter is repressed when TetO is expressed.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Description
PGK1-tetO is a PGK1 S.cerevisiae promoter with a tet-operon inserted 20 basepairs downstream of it’s TATA box(Figure 1). TetR has been shown to bind to and repress tetO containing synthetic promoter in S.cerevisiae[1]. TetO was placed downstrem of the TATA box in order to construct a promoter that was constitutive unless TetO was expressed in the same cell
Design
Characterization
Construct has been tested in an expression cassette containing PGK1-tetO upstream of a fluorescent mTurquoise gene, which simultaneously contained a tetR protein under the control of the blue light sensitive promoter C120-CYCp. In the presence of blue light and when transformed into a cell containing the appropriate factors to activate C120-CYCp, no difference was observed in the expression of mTurquoise(Figure 3) compared to the wildtype PGK1 promoter. This indicates that TetR did not successfully repress PGK1-TetO, and further engineering may be required to optimize this part.
