Part:BBa_K2290000

Fluoride Riboswitch regulated Chloramphenicol Acetyltranferase Operon

This is a composite part that allows for regulating the expression of a gene of interest with the B. ceres fluoride riboswitch. The functional parts of the switch are the following:

BBa_J23100 – a constitutively active promoter from the Anderson collection.

BBa_K2290008 – The B. cereus riboswitch with an engineered terminator that results in less read through/background. (This part is highly similar to part BBa_K911003 which is a part that includes both the Lys promoter and the riboswitch).

BBa_K2290004 – This is a sequence for a strong RBS that doesn't appear to be in the iGEM repository.

BBa_K2290005 – This is an E. coli optimized chloramphenicol resistance gene that we had synthesized.

BBa_B0010 – This is the T1 terminator from E. coli rrnB, reportedly a strong terminator

In theory, this part can be used to screen any transcriptional riboswitch or regulate the expression of any gene with the fluoride riboswitch.

is designed such that a user can easily clone in a new promoter and riboswitch or a gene of interest with Gibson cloning. To clone in a new promoter/riboswitch cut with HindIII. To clone in a new gene, cut with XhoI. Our part uses the E. coli constitutively active promoter J23100

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 31
  Illegal NheI site found at 54
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 182
  Illegal XhoI site found at 887
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]