Part:BBa_K2506004

USA_USP7 Promoter_USA

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 73
  Illegal AgeI site found at 1827
  
• 1000
  COMPATIBLE WITH RFC[1000]

Introduction

UUpromU is a system specailly designed for the SynNotch system (BBa_K2506001) that is capable of specifically expressing the target gene in the case of SynNotch. UUpromU consists of three parts: the promotor sequence of USP7 in the middle and three repeats of UAS gene in both 5’ and 3’ terminus. In order to activate the expression of USP7, we synthesized the UUpromU gene and the USP7 gene, and put it into pcDNA3.1 (+) plasmid (with the assistance from GenScript), where UUpromU replaced the CMV promoter in the pcDNA3.1(+). Therefore, in the case that the SynNotch system is stimulated by IL-17A, the Gal4-vp64 fusion protein that is detached from the cell membrane can specifically recognize and bind to the UAS sequence on UUpromU. Then, the transcription of USP7 gene can be motivated by VP-64, a transcription activator, which leads to the high expression of USP7. USP7 proteins can lead to de-ubiquitination of the FOXP3 protein, so that enhance the stability of FOXP3 protein in the inflammation environment by protecting FOXP3 from degradation via ubiquitination. Finally, the goal of regulating the stability of T cells can be achieved.