Part:BBa_K2353001:Design

tsPurpleDAS

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

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