DNA

Part:BBa_K5300042:Design

Designed by: Fan Yaxu   Group: iGEM24_CAU-China   (2024-09-30)


PglnK-sgRNA-GFP-tracrRNA-PnifH-gfp-pfa biosynthetic gene cluster


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 13428
    Illegal NheI site found at 17592
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2785
    Illegal BglII site found at 3698
    Illegal BglII site found at 7521
    Illegal BglII site found at 8671
    Illegal BglII site found at 9543
    Illegal BglII site found at 12800
    Illegal BglII site found at 15332
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2218
    Illegal NgoMIV site found at 2347
    Illegal NgoMIV site found at 2845
    Illegal NgoMIV site found at 4710
    Illegal NgoMIV site found at 5768
    Illegal NgoMIV site found at 6325
    Illegal NgoMIV site found at 6401
    Illegal AgeI site found at 3046
    Illegal AgeI site found at 4873
    Illegal AgeI site found at 7372
    Illegal AgeI site found at 8072
    Illegal AgeI site found at 10268
    Illegal AgeI site found at 10595
    Illegal AgeI site found at 13700
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1623
    Illegal SapI site found at 13529
    Illegal SapI site found at 15500
    Illegal SapI site found at 16700
    Illegal SapI site found at 18255
    Illegal SapI.rc site found at 10824
    Illegal SapI.rc site found at 11729
    Illegal SapI.rc site found at 19033


Design Notes

To ensure efficient expression of the pfa biosynthetic gene cluster in Sinorhizobium fredii CCBAU45436, we have added PPTase to the pfa biosynthetic gene cluster to increase production, at the same time, we have optimized the gene cluster's codon usage to suit the chassis.


Source

Sinorhizobium fredii CCBAU45436, Scytonema hofmannii, Aetherobacter fasciculatus SBSr002, Bradyrhizobium diazoefficiens USDA 110 (Bradyrhizobium japonicum USDA 110).

References