Part:BBa_K3271019:Design

His3::EX-RFP-SP pSB1K3 Type IIS Assembly

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found at 2291
  Illegal suffix found at 2347
  Illegal PstI site found at 2387
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2291
  Illegal SpeI site found at 2348
  Illegal PstI site found at 2362
  Illegal PstI site found at 2387
  Illegal NotI site found at 2297
  Illegal NotI site found at 2355
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2291
  Illegal XhoI site found at 1026
  Illegal XhoI site found at 2052
  Illegal XhoI site found at 2338
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 2291
  Illegal suffix found at 2348
  Illegal PstI site found at 2387
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 2291
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2306
  Illegal SpeI site found at 2348
  Illegal PstI site found at 2362
  Illegal PstI site found at 2387
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 2313
  Illegal BsaI.rc site found at 2341
  

Design Notes

Please note the upstream and downstream His3 locus homologies are found outside the prefix and suffix, not within the multiple cloning site. This contribution consists of an updated pSB1K3 BioBrick backbone that makes the BioBrick plasmids compatible with cloning in both E. coli and S. cerevisiae. This plasmid backbone is compatible with traditional and Type IIS cloning.

Source

The His3 upstream and downstream homologies are comprised of 100 bp each from the promoter region and the terminator region of the His3 locus of S. cerevisiae, respectively. The RFP cassette is identical in sequence to BBa_J04450.