Generator

Part:BBa_K4761203

Designed by: Pinhan Pan   Group: iGEM23_HiZJU-China   (2023-09-30)


T7-lacO-ARO10-adh6-T7-lacO-UGT85A1

Double expression rhodioloside generation unit. Controlled by lac operator. This part could be seen and constructed in pACYCDuet backbone.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1841
    Illegal SpeI site found at 792
    Illegal PstI site found at 670
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1841
    Illegal SpeI site found at 792
    Illegal PstI site found at 670
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1841
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1841
    Illegal SpeI site found at 792
    Illegal PstI site found at 670
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1841
    Illegal SpeI site found at 792
    Illegal PstI site found at 670
    Illegal NgoMIV site found at 2955
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

In 2023 HiZJU-China, this part is the salidroside generation unit installed on pACYCDuet(BBa_K4761103) as a double expression unit.

Fig 1 Characterization for successfully constructed plasmid pACYCDuet-ARO10-adh6-UGT85A1.

It is observed to increase salidroside yield compared with original fermentation strain pSA-ARO10+pET-UGT85A1 but on a limited level due to expression and metabolic stress caused.

Fig 2 HPLC results for (a)pSA-ARO10+pET-UGT85A1 and (b)pACYCDuet-ARO10-adh6-UGT85A1. Salidroside peak hard to observe due to byproducts from excessive UGT85A1 expression.
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