Part:BBa_K3179005

2Kt-E1A+E1B55K

This part is constructed by adding a 2X Kturn sequence before the E1A gene and E1B 55K of Ad5. Due to the existence of 2X Kturn, the translation of E1A and E1B 55K is regulated by L7Ae. E1A is an early gene of adenovirus and is essential in the commence of viral replication. E1B 55K can enhance the ability to lyse cells. In our design, we used P2A sequence to link E1A and E1B 55K, so that they can still maintain normal expression levels when using the same promoter. This part has an intron which also exists in E1A of the wild adenovirus. Using the splicing system in eukaryotes can make it function as normal. In our design, 2kt-E1A-E1B 55K is used as a response part to initiate the viral gene expression and lyse target cells.

We integrate 2kt-E1A-E1B 55K with miR885-5p-target and miR663b-target so that while miRNA885-5p and miRNA663b are absent in the cell, E1A can be translated and initiate viral gene expression to lyse target cells.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 859
  Illegal PstI site found at 1609
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1609
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2435
  Illegal BamHI site found at 63
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 859
  Illegal PstI site found at 1609
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 859
  Illegal PstI site found at 1609
  Illegal NgoMIV site found at 333
  Illegal NgoMIV site found at 1328
  Illegal AgeI site found at 69
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2461