DNA
Part:BBa_K4887005
Designed by: Duoqing LIN Group: iGEM23_Shanghai-BioX (2023-08-14)
Cas9
It is the gene of Cas9 (CRISPR-associated nuclease 9), a dual RNA-guided DNA endonuclease enzyme associated with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) adaptive immune system in Streptococcus pyogenes.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 907
Illegal PstI site found at 2329
Illegal PstI site found at 2533
Illegal PstI site found at 2563
Illegal PstI site found at 3775 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 907
Illegal PstI site found at 2329
Illegal PstI site found at 2533
Illegal PstI site found at 2563
Illegal PstI site found at 3775 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 368
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 907
Illegal PstI site found at 2329
Illegal PstI site found at 2533
Illegal PstI site found at 2563
Illegal PstI site found at 3775 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 907
Illegal PstI site found at 2329
Illegal PstI site found at 2533
Illegal PstI site found at 2563
Illegal PstI site found at 3775
Illegal NgoMIV site found at 1195
Illegal NgoMIV site found at 2299
Illegal NgoMIV site found at 2372
Illegal NgoMIV site found at 2857
Illegal NgoMIV site found at 3766
Illegal NgoMIV site found at 4227
Illegal NgoMIV site found at 4246 - 1000COMPATIBLE WITH RFC[1000]
5.5 Results:
(1) PCR detection for gene Cas9.
Genome DNA of the transgenic lines was extracted from leaves of these two regenerated seedlings. PCR detection on the genomes was performed by using primers designed based on the Cas9 protein gene. As the result, the transgenic lines 23216004 and 23216005 were further validated (Fig. 1).The sequences of the primers were as bellow:
- CAS9-F: 5’-atggactataaggaccacgacgg-3’
- CAS9-R: 5’-ttgtcgcctcccagctgagacag-3’
Fig. 1 Detection results of PCR for gene Cas9.
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Categories
Parameters
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