Difference between revisions of "Part:BBa K4046000"

Revision as of 20:11, 21 October 2021

FLAG-NLS-DhdR

This is a modified version of the transcriptional repression factor DhdR, which is responsive to the oncometabolite D-2-HG. DhdR is a transcriptional repression factor isolated from the bacteria Achromobacter denitrificans . The sequence was human codon optimized for expression in mammalian cells. Additionally, the sequence had a nuclear localization sequence attached to the end, for transport into the nucleus as part of our genetic reporter system, and a FLAG tag attached to the end.

A schematic for the operation of our system is shown below.

Figure 1: This figure outlines the mechanism of the interaction between the DhdR allosteric transcription factor and the dhdO binding site.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 675
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 310
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 186
Illegal BsaI site found at 303

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4046000 short</partinfo>
-This is a modified version of the transcriptional repression factor DhdR, which is responsive to the oncometabolite D-2-HG. The sequence was human codon optimized for expression in mammalian cells. Additionally, the sequence had a nuclear localization sequence attached to the beginning, for transport into the nucleus as part of our genetic reporter system, and a FLAG tag attached to the beginning, for ease of Western blot analysis.
+This is a modified version of the transcriptional repression factor DhdR, which is responsive to the oncometabolite D-2-HG. DhdR is a transcriptional repression factor isolated from the bacteria <i> Achromobacter denitrificans </i>. The sequence was human codon optimized for expression in mammalian cells. Additionally, the sequence had a nuclear localization sequence attached to the end, for transport into the nucleus as part of our genetic reporter system, and a FLAG tag attached to the end.
+A schematic for the operation of our system is shown below.
+[[File:T--Duke--DhdrSchematic.png|600 px]] <br>
+<b>Figure 1: This figure outlines the mechanism of the interaction between the DhdR allosteric transcription factor and the <i>dhdO</i> binding site.</b>