Difference between revisions of "Part:BBa K1926013"
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<partinfo>BBa_K1926013 short</partinfo> | <partinfo>BBa_K1926013 short</partinfo> | ||
This part is the mCHERRY UNIT of cyclebow system including a degradation tag named DBOX, a fluorescent gene mCherry and an sv40 terminator flanked by homodromous recognition site of recombinase VIKA named Vox. The whole part can be cut off by VIKA. | This part is the mCHERRY UNIT of cyclebow system including a degradation tag named DBOX, a fluorescent gene mCherry and an sv40 terminator flanked by homodromous recognition site of recombinase VIKA named Vox. The whole part can be cut off by VIKA. | ||
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| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K1926013 SequenceAndFeatures</partinfo> | ||
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| + | ===Usage=== | ||
You may use this part to: | You may use this part to: | ||
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1) Provide fluorescent red which can be quickly cut off by transient transfecting recombinase gene into nucleus; | 1) Provide fluorescent red which can be quickly cut off by transient transfecting recombinase gene into nucleus; | ||
| − | 2) Use it as a part of the flp-out system[ | + | 2) Use it as a part of the flp-out system[1]. |
5. Voutev, R. and E.J. Hubbard, A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans. Genetics, 2008. 180(1): p. 103-19. | 5. Voutev, R. and E.J. Hubbard, A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans. Genetics, 2008. 180(1): p. 103-19. | ||
| + | ===Source=== | ||
| − | + | The sequence of Vox and DBOX was retrieved from Addgene and got through denovo synthesis. mCHERRY and sv40 terminator were got from commercial plasmid: pentry dcas9 mcherry bira and pentry nls GFP/pcdna3.1, respectfully. | |
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| − | + | ===Construction Design=== | |
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| + | The mCherry with tags and RTSs are constructed using PCR and oligonucleotide ligation by the following procedure. Our UNITs all contain XbaI and SpeI enzyme sites, so we can easily line up the UNITs using 3A assembly. | ||
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| + | [[File:5-连接过程改.png|800px|thumb|left|'''Figure 1:''' Three steps to construct our UNITs using PCR and oligonucleotide ligation.]] | ||
| + | <br style="clear: both" /> | ||
Revision as of 11:54, 11 October 2016
The mCHERRY UNIT: mCherry flanked by Vox
This part is the mCHERRY UNIT of cyclebow system including a degradation tag named DBOX, a fluorescent gene mCherry and an sv40 terminator flanked by homodromous recognition site of recombinase VIKA named Vox. The whole part can be cut off by VIKA.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage
You may use this part to:
1) Provide fluorescent red which can be quickly cut off by transient transfecting recombinase gene into nucleus;
2) Use it as a part of the flp-out system[1].
5. Voutev, R. and E.J. Hubbard, A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans. Genetics, 2008. 180(1): p. 103-19.
Source
The sequence of Vox and DBOX was retrieved from Addgene and got through denovo synthesis. mCHERRY and sv40 terminator were got from commercial plasmid: pentry dcas9 mcherry bira and pentry nls GFP/pcdna3.1, respectfully.
Construction Design
The mCherry with tags and RTSs are constructed using PCR and oligonucleotide ligation by the following procedure. Our UNITs all contain XbaI and SpeI enzyme sites, so we can easily line up the UNITs using 3A assembly.
