Part:BBa_K678062

pJEJAM14

pJEJAM14 is a plasmid that after digestion with NotI is intended to be transformed into Aspergillus nidulans and integrated into the genome by non-homologous end joining. The plasmid consists the strong constitutive gpdA promoter , monomeric red fluorescent protein (mRFP1), the trpC terminator, a pyrG marker cassette for selection, an ori for propagation in E. coli, and an ampR resistance cassette (Figure 1).

DTU-Denmark-2 2011 Figure 1: Plasmid map of pJEJAM14, the figure is not drawn to scale.

When pJEJAM14 was integrated into A. nidulans expression of the gene encoding mRFP1, resulted in an even distribution of mRFP1 in the hyphae (Figure 2 and Figure 3). In figure 4 and figure 5 it be can see that the integration of the linearized pJEJAM14 must have impaired a gene or pathway of the transformant because its morphology and growth differed significantly from the [http://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild%20type wild-type].

DTU-Denmark-2 2011 Figure 2: Differential interference contrast image of Aspergillus nidulans with pJEJAM14.

DTU-Denmark-2 2011 Figure 2: Fluorescence image of Aspergillus nidulans with pJEJAM14.

DTU-Denmark-2 2011 Figure 2: Aspergillus nidulans Front

DTU-Denmark-2 2011 Figure 2: Aspergillus nidulans Back

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 3726
  Illegal XbaI site found at 5282
  Illegal XbaI site found at 8026
  Illegal PstI site found at 140
  Illegal PstI site found at 2657
  Illegal PstI site found at 4567
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 910
  Illegal PstI site found at 140
  Illegal PstI site found at 2657
  Illegal PstI site found at 4567
  Illegal NotI site found at 5239
  Illegal NotI site found at 8065
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 144
  Illegal BamHI site found at 3007
  Illegal BamHI site found at 4305
  Illegal XhoI site found at 700
  Illegal XhoI site found at 1082
  Illegal XhoI site found at 4165
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 3726
  Illegal XbaI site found at 5282
  Illegal XbaI site found at 8026
  Illegal PstI site found at 140
  Illegal PstI site found at 2657
  Illegal PstI site found at 4567
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 3726
  Illegal XbaI site found at 5282
  Illegal XbaI site found at 8026
  Illegal PstI site found at 140
  Illegal PstI site found at 2657
  Illegal PstI site found at 4567
  Illegal NgoMIV site found at 3643
  Illegal AgeI site found at 1289
  Illegal AgeI site found at 1437
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4358
  Illegal BsaI.rc site found at 1116
  Illegal BsaI.rc site found at 3357
  Illegal SapI site found at 322