Part:BBa_K5471020

pAAV-EF1a-mir195-eGFP-SV40pA

This plasmid construct is specially designed for the role of Transfer Gene plasmid in an rAAV2 production system, and for the plasmids replication in other cells.

This plasmid is comprised of the original pUC57 vector provided by Genscript, and the Cassette 2 https://parts.igem.org/Part:BBa_K5471013 cloned into position 2710.

Possesses all of the qualities of the Cassette mentioned above, which is the simoultaneous intragenic expression of mir195 and the strong expression of the fluorescent protein eGFP. The cassette, along with ITRs that flank it, is sure to make up the whole of the produced rAAV2 viral particle DNA, passing on its qualities to these viral vectors.

Possesses also all the qualities of the pUC57 vector, making this plasmid able to be massively replicated in other cells (eg. bacteria) and then be easily selected using Ampicillin.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1079
  Illegal XbaI site found at 1050
  Illegal PstI site found at 1022
  Illegal PstI site found at 3305
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1079
  Illegal NheI site found at 4854
  Illegal PstI site found at 1022
  Illegal PstI site found at 3305
  Illegal NotI site found at 4848
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1079
  Illegal BamHI site found at 1040
  Illegal XhoI site found at 3453
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1079
  Illegal XbaI site found at 1050
  Illegal PstI site found at 1022
  Illegal PstI site found at 3305
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1079
  Illegal XbaI site found at 1050
  Illegal PstI site found at 1022
  Illegal PstI site found at 3305
  Illegal NgoMIV site found at 3188
  Illegal AgeI site found at 2913
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2395
  Illegal BsaI site found at 3958
  Illegal SapI.rc site found at 767