Homing Endonuclease Design

I-GpeMI

Type

Wild-Type

Target Sequence

-11	-10	-9	-8	-7	-6	-5	-4	-3	-2	-1	1	2	3	4	5	6	7	8	9	10	11
T	T	T	C	C	G	C	T	T	A	T	T	C	A	A	C	C	C	T	T	T	A

FASTA Sequence

PTRNESINPWVLTGFADAEGSFILRIRNNNKSSAGYSTELGFQITLHKKDISILENIQSTWKVGVIANSGDNAVS
LKVTRFEDLRVVLNHFEKYPLITQKLGDYLLFKQAFSVMENKEHLKIEGIKRLVGIKANLNWGLTDELKEAFVAS
GGENIFVASGGERSLINKNIPNSGWLAGFTSGEGCFFVSLIKSKSKLGVQVQLVFSITQHARDRALMDNLVTYLG
CGYIKEKKKSEFSWLEFVVTKFSDIKDKIIPVFQVNNIIGVKLEDFEDWCKVAKLIEEKKHLTESGLEEIRNIKL
NMNKGRVL

Homology Model

PDB File

PyMOL File

Additional Information

Notes:

1.) Source organism:  Grosmannia penicillata  strain WIN(M)27

2.) Host gene:  Large subunit ribosomal RNA (rnl), mitochondrial

3.) Intron-encoded fusion protein

4.) Cleavage activity verified by flow cytometry via the tethered DNA cleavage assay (on the surface of yeast) and by an in vitro cleavage assay (with surface-released protein).  Orientation of target site is NOT experimentally verified, but strongly predicted through high homology to I-OnuI.

5.) Alternative sequence of endonuclease with mutations incorporated that are intended to increase overall solubility for in vitro studies:

PTRNESINPWVLTGFADAEGSFILRIRNNNKSSAGYSTELGFQITLHKKDKSILENIQSTWKVGVIANSGDNAVSLKVTRFEDLRVVLNHFEKYPLITQKLGDYLLFKQAFSVMENKEHLKIEGIKRLVGIKANLNWGLTDELKEAFVASGGENIFVASGGERSLINKNIPNSGWLAGFTSGEGCFFVSLIKSKSKLGVQVQLVFSITQHARDRELMDNLVTYLGCGYIKEKKKSEFSWLEFVVTKFSDIKDKIIPVFQENNIIGVKLEDFEDWCKVAKLIEEKKHLTESGLEEIRNIKLNMNKGRVL

6.) No structure

Date Added: 2011-09-30

LAHEDES: LAGLIDADG Homing Endonuclease Database and Engineering Server

Fred Hutchinson Cancer Research Center