Difference between revisions of "Part:BBa K3982000"

 
 
(2 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K3982000 short</partinfo>
 
<partinfo>BBa_K3982000 short</partinfo>
  
−
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 emits fluorescence.
+
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|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.

Cas14a1 gene with 6xHisTag and TEV site cloned in pSB1C3.jpeg

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE 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
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]