Part:BBa_K3271004

Ade2::KanMX-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. Between the upstream homology and the prefix is the KanMX selection marker which confers resistance to G418 in S. cerevisiae. 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 along with KanMX resistance 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 and plate on G418 selection.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found at 3820
  Illegal suffix found at 3842
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3820
  Illegal SpeI site found at 3843
  Illegal PstI site found at 3857
  Illegal NotI site found at 3826
  Illegal NotI site found at 3850
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3820
  Illegal XhoI site found at 1126
  Illegal XhoI site found at 2152
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 3820
  Illegal suffix found at 3843
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 3820
  Plasmid lacks a suffix.
  Illegal XbaI site found at 3835
  Illegal SpeI site found at 3843
  Illegal PstI site found at 3857
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.