![](https://parts.igem.org/images/partbypart/icon_coding.png)
Coding
Part:BBa_K4170005:Design
Designed by: Alexandros Giannopoulos Dimitriou Group: iGEM22_Thessaloniki_Meta (2022-09-27)
Lbucas13a assembly part 4
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal suffix found in sequence at 617
- 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 618
Illegal PstI site found at 632
Illegal NotI site found at 625 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 262
- 23INCOMPATIBLE WITH RFC[23]Illegal suffix found in sequence at 618
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 618
Illegal PstI site found at 632 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2
Illegal BsaI.rc site found at 640
Design Notes
This part derives from pGJK_His-SUMO-LbuCas13a plasmid (Plasmid #172488) by mutagenesis PCR to efficiently mutate illegal restriction sites which hinder compatibility with RFC [10] iGEM standard. In addition, the primers exploited incorporate BsaI recognition sites external of the amplified sequence to allow for efficient Golden Gate assembly with the parts 1-3 and the linearilized plasmid backbone.
Source
This part derives from pGJK_His-SUMO-LbuCas13a plasmid (Plasmid #172488)