Difference between revisions of "Part:BBa K2549041"

Revision as of 11:00, 9 October 2018

NLS-TEVp

This part is one of the downstream elements of our amplifier. It is constructed by fusing NLS and TEV protease, from N terminal to C terminal. NLS is a short nuclear location sequence from SV40 large T antigen. TEVp is a mutant TEV (tobacco etch virus) protease whose autoactivation is removed, thus performing more efficient cleavage. When coexpressed with VP64-dNLS-ZF21.16 in the same cell, it can destroy the nuclear location sequence and prevent the transcription activator from inducing the gene expression. Similarly, the cleavage can be conducted with the KRAB-dNLS-ZF21.16, thus keeping the repressor from switching off the gene expression.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 337

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 <partinfo>BBa_K2549041 short</partinfo>
-This part is one of the downstream elements of our amplifier. It is constructed by fusing NLS, TEVpN and CfaN, from N terminal to C terminal. NLS is a short nuclear location sequence from SV40 large T antigen. TEVpN is the N-terminal fragment of TEV protease. TEV protease is one of the best-characterized enzymes of the viral proteases which have more stringent sequence specificity. CfaN is the N-terminal fragment of Cfa which is a consensus sequence from an alignment of 73 naturally occurring DnaE inteins that are predicted to have fast splicing rates. When coexpressed with CfaC-TEVpC in the same cell, two fusion proteins can form and the site-specific cleavage can be realized.
+This part is one of the downstream elements of our amplifier. It is constructed by fusing NLS and TEV protease, from N terminal to C terminal. NLS is a short nuclear location sequence from SV40 large T antigen. TEVp is a mutant TEV (tobacco etch virus) protease whose autoactivation is removed, thus performing more efficient cleavage. When coexpressed with VP64-dNLS-ZF21.16 in the same cell, it can destroy the nuclear location sequence and prevent the transcription activator from inducing the gene expression. Similarly, the cleavage can be conducted with the KRAB-dNLS-ZF21.16, thus keeping the repressor from switching off the gene expression.
 <!-- Add more about the biology of this part here