Part:BBa_K758003

UAS, this part consists of five UAS sequences and hsp70 TATA.

This part includes five UAS(Upstream Activating Sequence) sequences, GAL4 binding sites. This part is working in mainly eukaryotic life.

Supplementary Experiment

Group: CSU_CHINA 2021

Author: Xingjun Zhao

UAS (BBa_K758003) is a piece of small DNA sequence that can be recognized and combined by GAL4 DBD. Team iGEM12_KIT-Kyoto repeated UAS 5 times when using it. In our design, considering of that the mutual effect of our blue light protein would reduce the efficiency and the whole pathway is relatively long, while the react time of our system should be as short as possible, we have made some alterations

We have repeated the UAS before promoter for 9 times. The purpose of constructing 9XUAS-Ad-LUC (BBa_K3734016) and 9XUAS-Ad-insulin (BBa_K3734033) is to enhance the recognition and combination of GAL4 DBD, improve the efficiency of downstream target gene activation.

Fig.1 Light controlled system testing experiment

Fig.2 Insulin concentration in supernatant (excluding insulin in medium) after blue light irradiation and dark treatment respectively

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 377
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 1000
  COMPATIBLE WITH RFC[1000]