Difference between revisions of "Part:BBa K4031000"

Revision as of 14:09, 21 October 2021

LbCpf1(Cas12a)

CRISPR-Cas12a (Cpf1) is a DNA endonuclease with RuvC as protein domain. Cas12a encoded by this part is derived from Lachnospiraceae bacterium ND2006 and recognizes and cleaves single- and double-stranded DNA sequences as targets. When Cas12a recognizes double-stranded DNA, it requires the PAM sequence of TTTN at the 5' end upstream of the opposite strand of the target sequence. Cas12a binds to target DNA upon gRNA induction and acquires non-specific ssDNA cleavage activity. To improve the convenience of BBa_K4031000 as a BioBrick, the recognition sites of restriction enzymes SapI and PstI in the coding sequence are deleted by silent mutation. In order not to affect the expression, we added silent mutation in consideration of the codon usage of E. coli.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1371
Illegal BglII site found at 2108
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 3022
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K4031000 short</partinfo>
-CRISPR-Cas12a (Cpf1) is a DNA endonuclease with RuvC as protein domain. Cas12a encoded by this part is derived from <i>Lachnospiraceae bacterium ND2006<i> and recognizes and cleaves single- and double-stranded DNA sequences as targets. When Cas12a recognizes double-stranded DNA, it requires the PAM sequence of TTTN at the 5' end upstream of the opposite strand of the target sequence. Cas12a binds to target DNA upon gRNA induction and acquires non-specific ssDNA cleavage activity.
+CRISPR-Cas12a (Cpf1) is a DNA endonuclease with RuvC as protein domain. Cas12a encoded by this part is derived from <i>Lachnospiraceae bacterium ND2006</i> and recognizes and cleaves single- and double-stranded DNA sequences as targets. When Cas12a recognizes double-stranded DNA, it requires the PAM sequence of TTTN at the 5' end upstream of the opposite strand of the target sequence. Cas12a binds to target DNA upon gRNA induction and acquires non-specific ssDNA cleavage activity.
-To improve the convenience of BBa_K4031000 as a BioBrick, the recognition sites of restriction enzymes SapI and PstI in the coding sequence are deleted by silent mutation. In order not to affect the expression, we added silent mutation in consideration of the codon usage of <i>E. coli<i>.
+To improve the convenience of BBa_K4031000 as a BioBrick, the recognition sites of restriction enzymes SapI and PstI in the coding sequence are deleted by silent mutation. In order not to affect the expression, we added silent mutation in consideration of the codon usage of <i>E. coli</i>.
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