Difference between revisions of "Part:BBa K2353000:Design"
(→‎Design Notes)
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tsPurple is intended to be assembled with pLambdaR-LacI (which is the promoter and regulatory system) and pLac-ClpXP-CI (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 tsPurple and LacI represses pLac, preventing transcription of ClpXP-CI. Upon induction with IPTG, IPTG binds to LacI, which prevents repression of pLac. Therefore, ClpXP-CI is transcribed; since there is no degradation tag attached, this should not change the levels of expression of tsPurple. The levels of expression of tsPurple with degradation tags can then be compared to the level of tsPurple expression (as a base-line comparison).
 
tsPurple is intended to be assembled with pLambdaR-LacI (which is the promoter and regulatory system) and pLac-ClpXP-CI (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 tsPurple and LacI represses pLac, preventing transcription of ClpXP-CI. Upon induction with IPTG, IPTG binds to LacI, which prevents repression of pLac. Therefore, ClpXP-CI is transcribed; since there is no degradation tag attached, this should not change the levels of expression of tsPurple. The levels of expression of tsPurple with degradation tags can then be compared to the level of tsPurple expression (as a base-line comparison).
 
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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).
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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
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===Source===
 
===Source===

Revision as of 14:52, 1 November 2017


tsPurple


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

tsPurple is intended to be assembled with pLambdaR-LacI (which is the promoter and regulatory system) and pLac-ClpXP-CI (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 tsPurple and LacI represses pLac, preventing transcription of ClpXP-CI. Upon induction with IPTG, IPTG binds to LacI, which prevents repression of pLac. Therefore, ClpXP-CI is transcribed; since there is no degradation tag attached, this should not change the levels of expression of tsPurple. The levels of expression of tsPurple with degradation tags can then be compared to the level of tsPurple expression (as a base-line comparison).
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

Source

This part is composed from sequences found in the parts registry.

References

Part:BBa_K1033906. (n.d.). Retrieved November 01, 2017, from https://parts.igem.org/wiki/index.php?title=Part%3ABBa_K1033906 Chromogenic Protein Paintbox™ - E. coli – pMOTHER vectors. (n.d.) Retrieved November 01, 2017, from https://www.atum.bio/eCommerce/catalog/datasheet/526
Eurofins Genomics. (n.d.) Retrieved November 01, 2017, from https://www.eurofins.com