Part:BBa_K3252033

gRNAJ23115-dCas9handle

Part BBa_K3252033 is the gRNA coding sequence that will express under the regulation of pQrr4 short promoter (BBa_K32252024). The gRNA will repress the BBa_J23115 promoter (BBa_K3252023) that we used for regulate the GFPmut3b (BBa_K3252031) expression as reporter gene. In our iGem team project this part has been assemble in BBa_K3252039 and BBa_K3252038 transcription unit with BBa_K3252035 transcription terminator (S. pyogenes terminator). This part also has been contain the dCas9 RNA handle that needed to build the dCas9-gRNA complex. That parts were assembled by order custom gene synthetic.

Characterization of this part was performed with the transcriptional unit BBa_K3252044. By using our experimental protocol, we characterize expression level which is regulated by this part in different level of present the AI consentration as inducer Lux cascade system.

Different level of AI consentration will influence the the level of expression of some gene. In this case expresssion level of GFPmut3b was observed to evaluate the repression level.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 47
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]