Composite

Part:BBa_K3809007

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


PuDHT + gldA homology regions

This part corresponds to the coding sequence of Paralcaligenes ureilyticus Dehydratase. It is regulated by BBa_J23100 promoter and has a BBa_B0030 RBS and an rrnB T1 terminator BBa_B0010. It also has a spacer between the promoter and the RBS (BBa_B0040). The sequence contains two homology regions for the gene glycerol dehydrogenase (gldA) of E. coli, which is a key enzyme in the glycerol methabolic pathway. We added this homology region in order to perform a knockout of gldA through phage plasmid pSIJ8. Instead of gldA, E. coli will express the gene of PuDHT, which codes for an enzyme that converts D-glycerate into pyruvate.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 380
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1123
    Illegal NgoMIV site found at 1336
    Illegal NgoMIV site found at 1744
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 407


[edit]
Categories
Parameters
None