Part:BBa_K3271000

Ade2::EXSP pSB1K3

The prefix and suffix (EXSP) of the pSB1K3 backbone are flanked by the upstream and downstream homology sequences of the Ade2 locus from S. cerevisiae, respectively. To target a DNA part of interest to the Ade2 locus in S. cerevisiae, insert the DNA part within the prefix and the suffix using any method of your choice. Then, digest the plasmid using PmeI. This creates a DNA fragment comprised of the DNA part of interest flanked by the Ade2 locus upstream homology and the downstream homology at the 5' and 3' ends of the part, respectively. Transform the digest into S. cerevisiae using a lithium acetate protocol of choice. Homologous recombination will facilitate the insertion of the DNA part of interest in the Ade2 locus.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found at 2276
  Illegal suffix found at 2298
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2276
  Illegal SpeI site found at 2299
  Illegal PstI site found at 2313
  Illegal NotI site found at 2282
  Illegal NotI site found at 2306
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2276
  Illegal XhoI site found at 1012
  Illegal XhoI site found at 2038
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 2276
  Illegal suffix found at 2299
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 2276
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2291
  Illegal SpeI site found at 2299
  Illegal PstI site found at 2313
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.