Part:BBa_K1392973

AracC+Antiholin+RBS+TetLVA+Term+TetR Promoter+ T4 Endolysin+Holin+Terminator

This part is a long Composite which contains many parts. Holin part comes from a Bacteriophage Holin Gene pS105 This is a variation of the S gene of the "Cell Lysis Cassette." When it is paired with an activator, the cassette is expressed and cell lysis is induced. The S105 gene has the initial 6 base pairs at the beginning of the wild type S gene sequence removed. This causes cell lysis to be induced at a faster rate than the wild type. The added Terminator is artificial and large. Composite part fot TetLVA TetPromoter and T4: This part is a composite structure which contains a promoter, RBS side and and a tetracycline repressor from transposon Tn10. It is transcribed in the opposite direction for the coding sequence of the repressor AraC. Through the binding to L+arabinose, the conformation of AraC changes.This causes the protein to diffuses from the DNA thereby inducing transcription. Through the RBS, the definition of the RBS efficiency could be observed. As the second part we added Terminator+Tetr Promoter+T4 Endolysin. For this part an artifical Terminator is used (generally added to another parts). The promoter is a TetR repressible promoter.It is sequenced for pTet inverting regulator. Promoter is constitutively ON and repressed by TetR. TetR repression is inhibited by the addition of tetracycline or its analog part

There are our graphs from modelling Kill-switch cycle under positive regulation and negative regulation.

In the first graph, you can see that without arabinose our promoter K808000 AraC Promoter doesn’t initiate then pTetR promoter R0040 initiates absence of the K808000 and produces antiholin so there isn’t any endolysin and holin activity in the cell metabolism.

However in the second graph; presence of arabinose, K808000 promoter initiates and cell start to produce endolysin and holin enzymes therefore cell autolyses itself after a while later.

Our parameter values present below in the table.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  Illegal AgeI site found at 2628
  Illegal AgeI site found at 2698
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961
  Illegal SapI site found at 1458