Difference between revisions of "Part:BBa K2353001:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | The promoter and terminator are both part of different parts, which means tspurple is not expressed unless it is assembled with pLambdaR-LacI | + | tsPurpleDAS is composed of a ribosomal binding site, tsPurple (a purple chromoprotein which functions as a reporter), and DAS (a moderately fast degradation tag). tsPurpleDAS is intended to be assembled with pLambdaR-LacI (BBa_K1911000),which is the promoter and regulatory system, and pLac-ClpXP-CI (BBa_K1911001),which is the non-lysosomal protease. The terminator is located at the beginning of pLac-ClpXP-CI. When fully assembled, p-lambda-r LacI transcribes tsPurpleDAS and LacI represses pLac, preventing transcription of ClpXP-CI. Upon induction with IPTG, LacI binds to IPTG which prevents repression of pLac. Therefore, ClpXP-CI is transcribed; ClpXP should recognize the DAS deg tag and degrade the tsPurple protein. The ensuing levels of expression could then be compared to that of BBa_K2353000 (tsPurple without a deg tag) to see the relative level of degradation experienced. |
| − | + | <br> | |
| − | + | IMPORTANT NOTE: The promoter and terminator are both part of different parts, which means tspurple is not expressed unless it is assembled with pLambdaR-LacI (BBa_K1911000). The protease complex of ClpX and ClpP is from part (BBa_K1911001) | |
| − | + | ||
===Source=== | ===Source=== | ||
| Line 18: | Line 17: | ||
===References=== | ===References=== | ||
Eurofins Genomics. (n.d.) Retrieved November 01, 2017, from https://www.eurofins.com | Eurofins Genomics. (n.d.) Retrieved November 01, 2017, from https://www.eurofins.com | ||
| − | Part:BBa_K1033906. (n.d.). Retrieved November 01, 2017, from https://parts.igem.org/wiki/index.php?title=Part%3ABBa_K1033906 | + | <br> |
| + | Chromogenic Protein Paintbox™ - E. coli – pMOTHER vectors. (n.d.) Retrieved November 01, 2017, from https://www.atum.bio/eCommerce/catalog/datasheet/526 | ||
| + | <br> | ||
| + | Part:BBa_K1033906. (n.d.). Retrieved November 01, 2017, from https://parts.igem.org/wiki/index.php?title=Part%3ABBa_K1033906 | ||
| + | <br> | ||
| + | Part:BBa_M0052. (n.d.) Retrieved November 01, 2017, from https://parts.igem.org/Part:BBa_M0052 | ||
Latest revision as of 16:09, 1 November 2017
tsPurpleDAS
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
tsPurpleDAS is composed of a ribosomal binding site, tsPurple (a purple chromoprotein which functions as a reporter), and DAS (a moderately fast degradation tag). tsPurpleDAS is intended to be assembled with pLambdaR-LacI (BBa_K1911000),which is the promoter and regulatory system, and pLac-ClpXP-CI (BBa_K1911001),which is the non-lysosomal protease. The terminator is located at the beginning of pLac-ClpXP-CI. When fully assembled, p-lambda-r LacI transcribes tsPurpleDAS and LacI represses pLac, preventing transcription of ClpXP-CI. Upon induction with IPTG, LacI binds to IPTG which prevents repression of pLac. Therefore, ClpXP-CI is transcribed; ClpXP should recognize the DAS deg tag and degrade the tsPurple protein. The ensuing levels of expression could then be compared to that of BBa_K2353000 (tsPurple without a deg tag) to see the relative level of degradation experienced.
IMPORTANT NOTE: The promoter and terminator are both part of different parts, which means tspurple is not expressed unless it is assembled with pLambdaR-LacI (BBa_K1911000). The protease complex of ClpX and ClpP is from part (BBa_K1911001)
Source
The sequences for this part came from the registry.
References
Eurofins Genomics. (n.d.) Retrieved November 01, 2017, from https://www.eurofins.com
Chromogenic Protein Paintbox™ - E. coli – pMOTHER vectors. (n.d.) Retrieved November 01, 2017, from https://www.atum.bio/eCommerce/catalog/datasheet/526
Part:BBa_K1033906. (n.d.). Retrieved November 01, 2017, from https://parts.igem.org/wiki/index.php?title=Part%3ABBa_K1033906
Part:BBa_M0052. (n.d.) Retrieved November 01, 2017, from https://parts.igem.org/Part:BBa_M0052