Coding

Part:BBa_K1484106

Designed by: Salil Bhate   Group: iGEM14_Cambridge-JIC   (2014-10-08)

LTI membrane localisation tag Localisation tag for plant reporters, which localises them to the cell membrane. Compatible with Plant Syntax (RFC105) as a C-Terminal fusion.

Usage and Biology

Add this tag to the 3' of your ORF. IMPORTANT: this contains flanking restriction sites for use with RFC105 compatible assembly.

When used in conjunction with fluorescent reporters, individual cell outlines are clearly visible with 5x magnification.

This tag can be very useful for counting cells, measuring cell size using automated computational techniques. See [Fernan Nature] for an illustration of how this tag can be used.

This tag is effective in many plant chassis:

In Marchantia, the fluorescent reporter appears as follows:

Simple routines can collect information about the sample cell sizes (in pixels).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 45
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 45
    Illegal NotI site found at 18
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 45
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 45
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 45
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 233


[edit]
Categories
//chassis/eukaryote/mpolymorpha
Parameters
None