Coding

Part:BBa_K4582000:Design

Designed by: Grégoire Leroy   Group: iGEM23_Aix-Marseille   (2023-10-08)


Coding sequence mucA* of P. putida MucA, resulting in alginate overproduction


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To enable homologous recombination after introducing the suicide plasmid into P. putida, it is necessary to add approximately 50-base pair sequences on both sides of the mutated mucA gene, which are homologous to the regions flanking the wild-type mucA gene on the chromosome. Subsequently, this entire construct is inserted into a suicide plasmid through cloning.



Source

We conducted a PCR on the wild-type strain P. putida KT2440 using a pair of primers, with the reverse primer hybridizing to the region where we intended to shorten the gene. Additionally, we added a stop codon to the 3' end of this primer.

References