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:
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.