Coding

Part:BBa_K3809006

Designed by: Francisco Javier Castañeda Villagran   Group: iGEM21_TecCEM   (2021-10-07)


PuDHT

This sequence codes for the enzyme dehydratase from the organism Paralcaligenes ureilyticus. It was designed as part of a novel selection method in which the enzyme converts D-glyceric acid into Pyruvate, which is a useful compound for bacteria. This gene will work alongside aldO and katE to develop a new selection mechanism which involves the knockout of genes gldA, glpK and oxyR to make E. coli suceptible to death by exposure to low concentrations of glycerol. We will substitute the genes gldA and glpK with PuDHT and aldO, and transform the bacteria with a plasmid containing selection cassette katE. AldO will convert glycerol to peroxide and D-glyceric acid, katE will convert peroxide to water and oxygen and PuDHT will convert D-glyceric acid into pyruvate.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 238
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 981
    Illegal NgoMIV site found at 1194
    Illegal NgoMIV site found at 1602
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 265


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Parameters
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