Coding

Part:BBa_K4392006:Design

Designed by: Atharva Valanju   Group: iGEM22_IISER-Pune-India   (2022-10-05)


ACC deaminase gene from Azospirillum lipoferum 4B


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 98
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 451
    Illegal NgoMIV site found at 478
    Illegal NgoMIV site found at 610
    Illegal NgoMIV site found at 975
    Illegal AgeI site found at 595
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 820


Design Notes

We worked with the pCZ-750 plasmid which can be mobilized into Azospirillum brasilense sp7. For cloning the gene we had to perform non-directional cloning using XbaI restriction sites, and so our primers were designed to amplify the acdS gene and add XbaI sites on both ends of the acdS gene.

Sequence of the primers:

acds:lipo:FP - GCTCTAGAATGCGCCTCGACCGTTTCG acds:lipo:RP - GCTCTAGATCAACCGTTGCGGTAGGTGTA



Source

This gene was amplified from the megaplasmid of Azosprillum lipoferum 4B

References