Part:BBa_K1300001

pTet promoter + Lock (crRNA) + ccdB + dterm

pTet is a constitutive promoter that is repressed by the presence of tetracycline. Without tetracycline, cis-repressed mRNA (crRNA) of ccdB (which is a toxin that inhibits the function of gyrase and interferes with helicase activity, thereby inhibiting processes such as DNA replication and RNA transcription)is continuously transcribed uninhibited. crRNA is an inactive form of mRNA because the cis-repressive sequence prevents protein expression from occurring. This is because the mRNA of the cis-sequence is complementary to the RBS in mRNA. As a result, a stem-loop formation occurs and blocks off the RBS so that ribosomes can no longer dock onto the mRNA.

If crRNA is considered a "locked" mRNA strand, a separate RNA sequence known as the trans-activating RNA (taRNA)(which is coded by part BBa_J01008) acts as a "key". taRNA binds onto the cis-repressive sequence and frees up the RBS so that protein expression can now occur. We have also designed a trans-activating sequence that is controlled by a promoter that responds to DNA damage that can be caused by UV radiation (BBa_K1300000)and a promoter that responds to UV radiation direction (BBa_K1300012) for the purpose of a kill switch system.

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 site found at 655