Part:BBa_K4675005:Design

pSB3CY_aeBlue shuttle vector backbone for Komagataella phaffii

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 4987
  Illegal SpeI site found at 2622
  Illegal PstI site found at 1105
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 4987
  Illegal NheI site found at 2681
  Illegal NheI site found at 2704
  Illegal NheI site found at 3673
  Illegal SpeI site found at 2622
  Illegal PstI site found at 1105
  Illegal NotI site found at 3614
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 4987
  Illegal BglII site found at 2518
  Illegal BamHI site found at 2441
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 4987
  Illegal SpeI site found at 2622
  Illegal PstI site found at 1105
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 4987
  Illegal SpeI site found at 2622
  Illegal PstI site found at 1105
  Illegal AgeI site found at 793
  Illegal AgeI site found at 905
  Illegal AgeI site found at 2083
  Illegal AgeI site found at 2406
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 1378
  Illegal BsaI site found at 4993
  Illegal BsaI.rc site found at 1099
  Illegal SapI site found at 1082
  Illegal SapI.rc site found at 6
  

Design Notes

We had to perform a mutagenesis of the PARS, as there was a SapI restriction site in this part.

Source

This shuttle vector was assembled using synthesized parts, iGEM biobricks (BBa_J435251) and PCR fragments. The plasmid from 2022 was used.