Part:BBa_K4720017

J23100+RBS+FleQ+TT+Pcl+Ppel+RBS

BBa_J23100 + RBS (BBa_J34801) FleQ (BBa_K4720000) + Double Terminator (BBa_B0015) + Pcl (BBa_R0051) + Ppel (BBa_K4242001) + RBS (BBa_J34801)

This part is the sensor part of our Cyclic-di-GMP biosensor design. Most of the parts design was obtained from CUG China 2022. A notable alteration is the replacement of the Pcl promoter in front of FleQ. This promoter was replaced by a constitutive promoter from the Anderson collection. The decision to alter this part was made later in our project due to the difficulty of obtaining this construct. It was hypothesized that recombination of the repeating Pcl sequence was causing the loss of the FleQ coding sequence. The part was ordered as two fragments of linear synthetic DNA. Three different approaches were made to assemble the construct: Gibson Assembly, Golden Gate with SapI and Golden Gate with BsaI. All attempts were however unsuccessful resulting in the absence of results.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 233
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]