Difference between revisions of "Part:BBa K4260008"
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| + | <strong><font size=5>Tryptophan, rho-independent terminator</font></strong></br> | ||
| + | <br><font size=3><strong>Type:</strong> Terminator</font></br> | ||
| + | <br><font size=3 color=red><strong>Designed by: </strong> -----</font></br> | ||
| + | <br><font size=3><strong>Group:</strong> iGEM_TecCEM</font></br> | ||
| + | <br> | ||
| + | <p align = "justify">This part is a 20 bp-long terminator, from the Trp operon, that forms a stem-loop structure. The tryptophan operator provides transcription efficiency as a terminator design, promoting a rho-independent or intrinsic termination. This means that the termination process carried out by Trp terminator does not depend on the presence of termination factors [1]. Moreover, it shares several features with other known sites of transcription termination of E. Coli. The tryptophan terminator was designed with scientific literature and iGEM team's research, in this way it was used for the transcription termination of a gene of another part (to know more, please visit <a href="https://parts.igem.org/Part:BBa_K729002">BBa_K4260006</a>), that was tested in E.coli DH5α and BL21 strains with experimental procedures. | ||
| + | </p> | ||
| + | <br><font size=3><strong><em>Sequence & Features</em></font></strong></br> | ||
| + | <br> | ||
| + | <center> | ||
| + | <img style="vertical-align: bottom;)" width=45% src="https://static.igem.org/mediawiki/parts/a/a5/K4260008_TecCem_Fig1.png"> | ||
| + | </center> | ||
| + | </br> | ||
| + | <center><strong>Fig. 1</strong> Sequence scheme.</center> | ||
| − | + | <br><strong>Length:</strong> 20 bp</br> | |
| − | + | <br><strong>Direction:</strong> Reverse</br> | |
| − | + | <br><strong>Sequence (5’ to 3’):</strong> CCCACTCAGTAGTGGGTTTT</br> | |
| − | + | <br> | |
| − | + | <p align = "justify">The secondary structure prediction was obtained with UNAFold, indicating a ΔG value of -5.20. Other data obtained from the model is shown in the table below.</p></br> | |
| − | < | + | <center> |
| − | + | <table> | |
| − | < | + | <tr> |
| − | + | <td><img style="vertical-align: bottom;)" width="300" height="400" src="https://static.igem.org/mediawiki/parts/b/b5/TrpTerm--TecCEM--.png"></td> | |
| − | + | <td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td> | |
| − | + | <td><img style="vertical-align: bottom;)" width="350" height="300" src="https://static.igem.org/mediawiki/parts/9/97/TablaTerminadorTrp.png"></td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td><center><strong>Fig. 2</strong>Structure prediction of the <br> Tpr terminator. [Obtained with UNAFold].</center></td> | |
| − | + | <td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td> | |
| + | <td><center><strong>Fig. 3</strong> Additional thermodynamic information of <br>the structure prediction. [Obtained with UNAFold]</center></td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | </center> | ||
| + | <br><font size=3><strong>References</font></strong></br> | ||
| + | <br> | ||
| + | <p align = "justify">[1] Chen, J., Morita, T. & Gottesman, S. (2019). <em>Regulation of Trsncription Termination of Small RNAs and by Smll RNAs: Molecular Mechanisims and Biological Functions. </em> Frontiers in Cellular and Infection Microbiology. <a href="https://doi.org/10.3389/fcimb.2019.00201">https://doi.org/10.3389/fcimb.2019.00201</a></p></br> | ||
<hr> | <hr> | ||
| − | + | <br> | |
| − | + | <p align = "justify">Tested to recognize its functionality in Thus experimental measuring was performed on the Safety project proposal by Tec CEM 2022 and it was found not to be difficult to use and has an automatic adjustment to experimental protocols. | |
| + | </p></br> | ||
| + | </html> | ||
Revision as of 01:12, 10 October 2022
Tryptophan, rho-independent terminator
Type: Terminator
Designed by: -----
Group: iGEM_TecCEM
This part is a 20 bp-long terminator, from the Trp operon, that forms a stem-loop structure. The tryptophan operator provides transcription efficiency as a terminator design, promoting a rho-independent or intrinsic termination. This means that the termination process carried out by Trp terminator does not depend on the presence of termination factors [1]. Moreover, it shares several features with other known sites of transcription termination of E. Coli. The tryptophan terminator was designed with scientific literature and iGEM team's research, in this way it was used for the transcription termination of a gene of another part (to know more, please visit BBa_K4260006), that was tested in E.coli DH5α and BL21 strains with experimental procedures.
Sequence & Features
Length: 20 bp
Direction: Reverse
Sequence (5’ to 3’): CCCACTCAGTAGTGGGTTTT
The secondary structure prediction was obtained with UNAFold, indicating a ΔG value of -5.20. Other data obtained from the model is shown in the table below.
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Tpr terminator. [Obtained with UNAFold]. |
the structure prediction. [Obtained with UNAFold] |
References
[1] Chen, J., Morita, T. & Gottesman, S. (2019). Regulation of Trsncription Termination of Small RNAs and by Smll RNAs: Molecular Mechanisims and Biological Functions. Frontiers in Cellular and Infection Microbiology. https://doi.org/10.3389/fcimb.2019.00201
Tested to recognize its functionality in Thus experimental measuring was performed on the Safety project proposal by Tec CEM 2022 and it was found not to be difficult to use and has an automatic adjustment to experimental protocols.