Difference between revisions of "Part:BBa K1926012"
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<partinfo>BBa_K1926012 short</partinfo> | <partinfo>BBa_K1926012 short</partinfo> | ||
| − | This part is the VIKA UNIT of cyclebow system including an sv40 nuclear location sequence (NLS), a recombinase gene named VIKA 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 improvement of the BBa_K1641003 | + | This part is the VIKA UNIT of cyclebow system including an sv40 nuclear location sequence (NLS), a recombinase gene named VIKA 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 improvement of the [[Part:BBa_K1641003]], which is a plain VIKA. |
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| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K1926012 SequenceAndFeatures</partinfo> | ||
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| + | ===Usage=== | ||
You may use this part to: | You may use this part to: | ||
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| − | + | ===Source=== | |
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| − | + | The recombinase gene VIKA and its RTS Vox were got through denovo synthesis. NLS and sv40 terminator were got from commercial plasmid: pentry nls GFP/pcdn3.1. | |
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| − | < | + | |
| + | ===Construction Design=== | ||
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| + | The VIKA 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|600px|thumb|left|'''Figure 1:''' Three steps to construct our UNITs using PCR and oligonucleotide ligation.]] | ||
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| + | ===Gel analysis after EcoR1 digestion=== | ||
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| + | The length of our biobricks are confirmed by gel analysis after EcoR1 digestion. The biobrick BBa_R0040, which is the negative control in 2016’s interlab, was used as a control because it only have a 54 bp part. | ||
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| + | [[File: 2016--SYSU-CHINA--biobrick2.png|400px|thumb|left|'''Figure 2:''' AGE image of biobricks BBa_K1926001-BBa_K1926003. The samples were pre dyed with gelred]] | ||
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Latest revision as of 07:29, 19 October 2016
The VIKA UNIT: recombinase VIKA flanked by Vox
This part is the VIKA UNIT of cyclebow system including an sv40 nuclear location sequence (NLS), a recombinase gene named VIKA 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 improvement of the Part:BBa_K1641003, which is a plain 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) Test the cut-off efficiency of recombinase VIKA by qRT-PCR;
2) Fast cut off this part by transient transfecting recombinase gene into nucleus;
3) Do lineage tracking.
Source
The recombinase gene VIKA and its RTS Vox were got through denovo synthesis. NLS and sv40 terminator were got from commercial plasmid: pentry nls GFP/pcdn3.1.
Construction Design
The VIKA 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.
Gel analysis after EcoR1 digestion
The length of our biobricks are confirmed by gel analysis after EcoR1 digestion. The biobrick BBa_R0040, which is the negative control in 2016’s interlab, was used as a control because it only have a 54 bp part.
