Part:BBa_K1214012

pScr-RBS-GFP

Sucrose operon promoter with GFP coding sequence downstream. Should constitutively express GFP. Regulated by ScrR (BBa_K1214005)

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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 765
  

Biology

The ScrR gene encodes a protein which normally binds to and represses pScr promoter. When sucrose is present in the system, it binds to the ScrR protein and prevents it from repressing the operon promoter.

"http://i826.photobucket.com/albums/zz188/lana_khazma/fancy%20stuff/sucrstuff_zps30aefeaf.png"

Testing

Promoter Strength

Promotion was tested against pFru (BBa_K1214007) and a constitutive promoter (BBa_K823014) using Flow cytometry. All three promoters were ligated in front of RBS-GFP to constitutively express GFP. The cells were inoculated for 18 hours at 37 C in 3 mL of chloramphenicol broth and then examined under fluorescent microscopy.


Flow cytometry was then completed to determine Mean GFP.
This shows that the pScr promoter strength is comparable to the well-characterized BBa_K823014 promoter.