Difference between revisions of "Part:BBa K3982000"
Akankshya16 (Talk | contribs) |
Akankshya16 (Talk | contribs) |
||
| Line 5: | Line 5: | ||
Cas14a1 is a type V-F exceptionally compact RNA-guided nuclease. Cas14 forms a complex with RNA-guided nuclease and it can cleave ssDNA without PAM specificity. It comes tagged with 6xHis and has the Tobacco Etch Virus (TEV) protease and cleavage site for easy purification. In our project CODE M, Cas14a1 and sgRNA complex is used to detect SNPs in the Mycobacterium tuberculosis genome in a sputum sample. Upon detection, it triggers cleavage of a Fluorescence-Quencher pair ssDNA reporter and thus fluorescence is emitted which confirms the presence of SNPs in the genome. | Cas14a1 is a type V-F exceptionally compact RNA-guided nuclease. Cas14 forms a complex with RNA-guided nuclease and it can cleave ssDNA without PAM specificity. It comes tagged with 6xHis and has the Tobacco Etch Virus (TEV) protease and cleavage site for easy purification. In our project CODE M, Cas14a1 and sgRNA complex is used to detect SNPs in the Mycobacterium tuberculosis genome in a sputum sample. Upon detection, it triggers cleavage of a Fluorescence-Quencher pair ssDNA reporter and thus fluorescence is emitted which confirms the presence of SNPs in the genome. | ||
| − | [[File:Cas14a1 gene with 6xHisTag and TEV site cloned in pSB1C3.jpeg| | + | [[File:Cas14a1 gene with 6xHisTag and TEV site cloned in pSB1C3.jpeg|700px|]] |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Latest revision as of 16:09, 1 October 2021
6xHis - Tagged Cas14a1 with TEV site
Cas14a1 is a type V-F exceptionally compact RNA-guided nuclease. Cas14 forms a complex with RNA-guided nuclease and it can cleave ssDNA without PAM specificity. It comes tagged with 6xHis and has the Tobacco Etch Virus (TEV) protease and cleavage site for easy purification. In our project CODE M, Cas14a1 and sgRNA complex is used to detect SNPs in the Mycobacterium tuberculosis genome in a sputum sample. Upon detection, it triggers cleavage of a Fluorescence-Quencher pair ssDNA reporter and thus fluorescence is emitted which confirms the presence of SNPs in the genome.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2047
Illegal EcoRI site found at 3487
Illegal XbaI site found at 2062
Illegal PstI site found at 14
Illegal PstI site found at 2277
Illegal PstI site found at 2443 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2047
Illegal EcoRI site found at 3487
Illegal PstI site found at 14
Illegal PstI site found at 2277
Illegal PstI site found at 2443
Illegal NotI site found at 7
Illegal NotI site found at 2053 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2047
Illegal EcoRI site found at 3487
Illegal BamHI site found at 3081
Illegal XhoI site found at 1031
Illegal XhoI site found at 1923 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 2047
Illegal EcoRI site found at 3487
Illegal PstI site found at 14
Illegal PstI site found at 2277
Illegal PstI site found at 2443 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 2047
Illegal EcoRI site found at 3487
Illegal XbaI site found at 2062
Illegal PstI site found at 14
Illegal PstI site found at 2277
Illegal PstI site found at 2443
Illegal AgeI site found at 2737 - 1000COMPATIBLE WITH RFC[1000]