Difference between revisions of "Part:BBa K5034222:Design"
 
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===Design Notes===
 
===Design Notes===
  
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PPK1 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 <i>E. coli</i> DH5&#945; to amplify, and was purified by picking monoclonal culture and sequencing.
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PPK1 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator(after EcoRI and XhoI digestion) to get the expression vector. It then got transformed into <i>E. coli</i> DH5&#945; to amplify, and was purified by picking monoclonal culture and sequencing.
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The terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201).
  
 
===Source===
 
===Source===

Latest revision as of 11:24, 30 September 2024


Pi <-> Poly P


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

PPK1 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator(after EcoRI 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.

The terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201).

Source

Polyphosphate kinase 1(PPK1) from Citrobacter freundii. NBCI reference sequence: NZ_LR890181.1:c1367205-1365139

References

Wang, X., Wang, X., Hui, K., Wei, W., Zhang, W., Miao, A., . . . Yang, L. (2018). Highly Effective Polyphosphate Synthesis, Phosphate Removal, and Concentration Using Engineered Environmental Bacteria Based on a Simple Solo Medium-Copy Plasmid Strategy. Environmental Science & Technology, 52(1), 214-222. doi:10.1021/acs.est.7b04532