Composite

Part:BBa_K4621133:Design

Designed by: Anyang Chen, Ruikang Chen   Group: iGEM23_SCUT-China   (2023-10-05)


The composite Parts of SCUT-3-GABAi9 that achieve high GABA production functiont.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 5966
    Illegal PstI site found at 3860
    Illegal PstI site found at 4094
    Illegal PstI site found at 5306
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 5943
    Illegal SpeI site found at 5966
    Illegal PstI site found at 3860
    Illegal PstI site found at 4094
    Illegal PstI site found at 5306
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1899
    Illegal BglII site found at 2694
    Illegal BglII site found at 5463
    Illegal BamHI site found at 1639
    Illegal BamHI site found at 2988
    Illegal BamHI site found at 5931
    Illegal BamHI site found at 6136
    Illegal XhoI site found at 2312
    Illegal XhoI site found at 4658
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 5966
    Illegal PstI site found at 3860
    Illegal PstI site found at 4094
    Illegal PstI site found at 5306
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 5966
    Illegal PstI site found at 3860
    Illegal PstI site found at 4094
    Illegal PstI site found at 5306
    Illegal NgoMIV site found at 53
    Illegal NgoMIV site found at 1242
    Illegal NgoMIV site found at 3256
    Illegal NgoMIV site found at 3627
    Illegal NgoMIV site found at 3830
    Illegal AgeI site found at 470
    Illegal AgeI site found at 690
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4838
    Illegal BsaI.rc site found at 5717
    Illegal SapI site found at 2230
    Illegal SapI site found at 3520
    Illegal SapI.rc site found at 4717


Design Notes

The effectiveness, and GC content.


Source

The CRISPRi fragment is derived from PZ-gadB-i9 plasmid, then be ligated after EctOperon on the PZ-gadB-i9 plasmid.

References

[1] Guo, X. F., Hagiwara, T., Masuda, K., and Watabe, S. (2011) Biosci. Biotechnol. Biochem. 75, 1867-1871. [2] Zhao Y, Li L, Zheng G, Jiang W, Deng Z, Wang Z, Lu Y. CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces. Biotechnol J. 2018 Sep;13(9):e1800121. doi: 10.1002/biot.201800121. Epub 2018 Jul 4. PMID: 29862648.