Part:BBa_K5034231:Design

Poly P -> Poly P(smaller) or Pi

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 11
  Illegal PstI site found at 3787
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 11
  Illegal PstI site found at 3787
  Illegal NotI site found at 2828
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 11
  Illegal BglII site found at 3574
  Illegal XhoI site found at 4985
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 11
  Illegal PstI site found at 3787
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 11
  Illegal PstI site found at 3787
  Illegal NgoMIV site found at 556
  Illegal NgoMIV site found at 4238
  Illegal NgoMIV site found at 4521
  Illegal AgeI site found at 396
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3150
  Illegal SapI.rc site found at 4087
  Illegal SapI.rc site found at 4297
  

Design Notes

PPN1 gene was PCR-amplified and inserted into the plasmid pYYDT(after NdeI and XhoI digestion) to get the expression vector. It then got transformed into E. coli DH5α to amplify, and was purified by picking monoclonal culture and sequencing.

Source

Endopolyphosphatase(PPN1) from Saccharomyces cerevisiae. NCBI reference sequence: NC_001136.10.

References

Lichko, L. P., Kulakovskaya, T. V., & Kulaev, I. S. (2006). Inorganic polyphosphate and exopolyphosphatase in the nuclei of Saccharomyces cerevisiae: dependence on the growth phase and inactivation of the PPX1 and PPN1 genes. Yeast, 23(10), 735-740. doi:10.1002/yea.1391