Part:BBa_K4898000
Inducible dual promoter system
Background
This is a inducible dual promoter system incorporated in a vector. First is the Ptac, short for the Tac-Promoter, is a synthetic DNA regulatory element widely used in molecular biology and genetic engineering. It is engineered by merging components of the promoters found in the trp and lac operons, two important genetic systems in bacteria. Ptac is a valuable tool for controlling gene expression in various experimental settings, allowing researchers to precisely modulate the transcription of specific genes. Its versatility makes it a cornerstone in the construction of genetically modified organisms and the production of recombinant proteins, enabling a wide range of biotechnological applications. It can be induced by IPTG. The other is the Ptet, or the Tet-Promoter. This promoter is regulated by the presence or absence of tetracycline or its analogs, providing researchers with a powerful tool for controlling gene transcription. In the absence of tetracycline, the Tet promoter is active, allowing gene expression. When tetracycline or its derivatives are introduced, they bind to a repressor protein, which then prevents the promoter from initiating transcription. This unique on-off switch-like mechanism makes the Tet promoter an invaluable asset in genetic studies, enabling precise regulation of gene expression in a variety of experimental and biotechnological contexts. Overall this provides a powerful tool for wide variety of applications in gene expression.
Part Uses
1. Expressing a protein: A protein with its chaperone can be cloned under each of the promoters allowing tight control and regulation for folding in protein production. This may be used to prevent metabolic burden on the cells to produce chaperones.
2. When expressing different subunits of a protein: This can have the coding sequences of two subunits under each promoter for tight regulation.
3. As a kill switch: For example incorporating the ccdb toxin anti-toxin gene under the Ptet promoter (well controlled using doxycycline) when induced will act as a kill switch for the GMOs.
4. In-vivo removal of fusion tags: Under the Ptac promoter gene responsible for protein synthesis with the fusion tag can be incorporated while a protease specific to the fusion tag may be added under the Ptet promoter allowing the tag to be cleaved when desired through induction.
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]
| None |