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Revision as of 17:02, 13 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 BBa_K1641003, which is a plain VIKA. --[1]

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
    COMPATIBLE 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.

Figure 1: Three steps to construct our UNITs using PCR and oligonucleotide ligation.