Part:BBa_K678050

pJEJAM2

pJEJAM2 is a plasmid intended for transient transfection of mammalian cells. pJEJAM2 consists of the strong constitutive cytomegalovirus (CMV) promoter, green fluorescence protein (GFP) with the peroxisomal targeting sequence 1 (PTS1) fused to the C-terminal, the BGK polyA, the hygR marker cassette, an ori for propagation in E.coli and, a ampR resistance cassette (Figure 1).

DTU-Denmark-2 2011 Figure 1: Plasmid map of pJEJAM2, the figure is not drawn to scale.

PTS1 with the amino acid sequence S-K-L should ensure the localization of GFP to the peroxisomes of the cell. Although the microscopy shows GFP distributed in points that we assume are the peroxisomes, further analysis are necessary to confirm that GFP is in fact is targeted to the peroxisomes (Figure 2).

DTU-Denmark-2 2011 Figure 2: U-2 OS cells transiently transfected with pJEJAM2 expressing GFP localized to the peroxisomes. As can be seen on the picture this vector most likely localizes GFP to the peroxisomes of the cells. The white bar has a length of 30μm.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2225
  Illegal PstI site found at 2315
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2225
  Illegal PstI site found at 2315
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2225
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 2225
  Illegal PstI site found at 2315
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 2225
  Illegal PstI site found at 2315
  Illegal NgoMIV site found at 3080
  
• 1000
  COMPATIBLE WITH RFC[1000]