Difference between revisions of "Part:BBa K2066113"
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<partinfo>BBa_K2066113 short</partinfo> | <partinfo>BBa_K2066113 short</partinfo> | ||
| − | The synthetic enhancer, first characterized by Amit et. al. 2011 allows for a multistate transfer function by modulating the rigidity of the spacer region between the enhancer promoter region, thus affecting the DNA rigidity and hence the kinetic capability of the two components to loop together and initiate transcription. | + | The synthetic enhancer, first characterized by Amit et. al. 2011, allows for a multistate transfer function by modulating the rigidity of the spacer region between the enhancer and promoter region, thus affecting the DNA rigidity and hence the kinetic capability of the two components to loop together and initiate transcription. For more information on the mechanism, please refer to: http://2016.igem.org/Team:William_and_Mary/Synthetic_Enhancer. |
This synthetic enhancer part consists of a 2x TetO binding cassette within the spacer region between the enhancer and promoter. The two binding sites allows for three discrete states of output: a repressed, intermediate, and unrepressed state. Depending on the steady state of TetR, the amount of bound TetR varies and thus affects the rigidity of the spacer region as well as the kinetics of the looping between the enhancer and promoter, which allows for the initiation of transcription. This part has to be transformed in conjugation with the pACT Tet plasmid (Bba_K2066037), which controls the expression of TetR and NRII2302, a kinase that activates the NRI product (which binds and primes the upstream enhancer region). | This synthetic enhancer part consists of a 2x TetO binding cassette within the spacer region between the enhancer and promoter. The two binding sites allows for three discrete states of output: a repressed, intermediate, and unrepressed state. Depending on the steady state of TetR, the amount of bound TetR varies and thus affects the rigidity of the spacer region as well as the kinetics of the looping between the enhancer and promoter, which allows for the initiation of transcription. This part has to be transformed in conjugation with the pACT Tet plasmid (Bba_K2066037), which controls the expression of TetR and NRII2302, a kinase that activates the NRI product (which binds and primes the upstream enhancer region). | ||
Latest revision as of 05:29, 29 October 2016
Synthetic Enhancer with 2X TetO cassette (55as) on UNS backbone
The synthetic enhancer, first characterized by Amit et. al. 2011, allows for a multistate transfer function by modulating the rigidity of the spacer region between the enhancer and promoter region, thus affecting the DNA rigidity and hence the kinetic capability of the two components to loop together and initiate transcription. For more information on the mechanism, please refer to: http://2016.igem.org/Team:William_and_Mary/Synthetic_Enhancer.
This synthetic enhancer part consists of a 2x TetO binding cassette within the spacer region between the enhancer and promoter. The two binding sites allows for three discrete states of output: a repressed, intermediate, and unrepressed state. Depending on the steady state of TetR, the amount of bound TetR varies and thus affects the rigidity of the spacer region as well as the kinetics of the looping between the enhancer and promoter, which allows for the initiation of transcription. This part has to be transformed in conjugation with the pACT Tet plasmid (Bba_K2066037), which controls the expression of TetR and NRII2302, a kinase that activates the NRI product (which binds and primes the upstream enhancer region).
Source: The enhancer, tet cassette (55as), glnAp2 synthetic promoter, NRI coding region, and mCherry coding region sequences were derived from Amit, R., Garcia, H. G., Phillips, R. & Fraser, S. E. Building enhancers from the ground up: a synthetic biology approach. Cell146, 105–118 (2011). The UNS sequences at the ends of the insert are derived from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860. A huge thanks to all the researchers involved in its original creation!
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 111
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 890
- 1000COMPATIBLE WITH RFC[1000]