Part:BBa_K1300003
Constitutive T7 promoter + Lock (crRNA) + ccdB + dterm
The T7 promoter is a constitutive promoter that produces high levels of transcription in E. coli. As a result, this part will constantly be producing 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). 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)cfor the purpose of a kill switch system. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 315
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