Plasmid

Part:BBa_K4231018:Design

Designed by: Shi Han   Group: iGEM22_USTC   (2022-09-30)
Revision as of 02:54, 1 October 2022 by Registry (Talk | contribs)

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pUC19 carrying with IDI, NADH-HMG, ERG8, ERG19 and URA3 tag from Y.lipolytica


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2325
    Illegal EcoRI site found at 8712
    Illegal PstI site found at 862
    Illegal PstI site found at 1992
    Illegal PstI site found at 7315
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2325
    Illegal EcoRI site found at 8712
    Illegal NheI site found at 634
    Illegal PstI site found at 862
    Illegal PstI site found at 1992
    Illegal PstI site found at 7315
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2325
    Illegal EcoRI site found at 8712
    Illegal BglII site found at 719
    Illegal BglII site found at 6238
    Illegal BamHI site found at 974
    Illegal BamHI site found at 1073
    Illegal BamHI site found at 1613
    Illegal BamHI site found at 6936
    Illegal BamHI site found at 8102
    Illegal XhoI site found at 3028
    Illegal XhoI site found at 3862
    Illegal XhoI site found at 3895
    Illegal XhoI site found at 6653
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2325
    Illegal EcoRI site found at 8712
    Illegal PstI site found at 862
    Illegal PstI site found at 1992
    Illegal PstI site found at 7315
  • 25
    INCOMPATIBLE WITH RFC[25]
    Unknown
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 7991
    Illegal BsaI.rc site found at 2386
    Illegal BsaI.rc site found at 3048
    Illegal BsaI.rc site found at 3396
    Illegal BsaI.rc site found at 7710
    Illegal BsaI.rc site found at 8060


Design Notes

All parts were combined by restriction enzyme, T4 DNA ligase and infusion cloning.Considering homologous recombination in Y.lipolytica, we made homologous sequences pTef1 and pGAP as far apart as possible.


Source

The coding sequences of IDI, ERG8, ERG19, URA3 were amplified from Y.lipolytica.The ORF of NADH-HMG was found in NCBI and was synthesized according to the sequence.

References