Part:BBa_K2970004

Trigger 2 Composition

Trigger 2 forms together with Trigger 1 (BBa_K2970000) a complex due to complementary regions, as shown in Figure 1A. This complex can open a gate (BBa_K2970002, Figure 1B) to enable translation of a gene that is locked with this gate to regulate translation. After binding of the trigger complex due to complementarity, the hairpin of the gate will open and release the ribosome binding site and the start codon, thus translation can occur (Figure 2). The affinity between the trigger complex and the gate is greater than that of the gate to itself (in the loop). A single trigger cannot open the gate because it has only half the required complementary sequence.

Figure 1: A) Formation of trigger complex after translation. B) mRNA of gate sequence forms secondary structures that hide the ribosome binding site and the start codon.

Figure 2: Opening of the gate due to annealing of trigger complex to gate.

We used this system to regulate a chloramphenicol resistance, which is locked by the gate. To transform the trigger into bacteria it should be put on a plasmid together with a promoter and a terminator as we did with this part. After transformation of both trigger plasmids (BBa_K2970003 and BBa_K2970004) and the gate plasmid (BBa_K2970006) in one bacterium all three mRNA structures will be formed, the gate will be opened, and the translation of the chloramphenicol resistance can take place. Bacteria that took all three plasmids are able to survive on media with chloramphenicol. This system can be used to transform many genes of interest on three different plasmids into bacteria with only using one antibiotic resistance instead of three different resistances.

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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]