Protein_Domain
Fok_i

Part:BBa_K243001:Design

Designed by: Freiburg Bioware09   Group: iGEM09_Freiburg_bioware   (2009-10-08)
Revision as of 14:55, 8 October 2009 by Timo (Talk | contribs) (→‎References)

Protein domain (inactive) of the restriction endonuclease FokI


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]


Design Notes

Modifications of the vector (catalytic inactive heterodimer) -heterodimeric amino acids 

  * switch Glutamin 486/298-300 to Glutamate (CAA->GAA)

  * switch Isoleucin 499/337-339 to Leucin (ATC->CTG)

-catalytic amino acids 

  * switch Aspartate 450/190-192 to Alanin (GAC->GCG)

  * switch Aspartate 467/243-245 to Alanin (GAT->GCG)


Source

extract coding region of Fok from the restriction-modification genes of the chromosomal DNA of Flavobacterium okeanokoites. Part synthesized by Mr.Gene

References

Mary C. Looneya, Laurie S. Morana, William E. Jacka, George R. Feeherya, Jack S. Bennera, Barton E. Slatkoa and Geoffrey G. Wilson;(1989)
Nucleotide sequence of the FokI restriction-modification system: separate strand-specificity domains in the methyltransferase; Gene Vol.80 Issue:2 Pages:193-208


Jeffrey C Miller1, Michael C Holmes1, Jianbin Wang1, Dmitry Y Guschin1, Ya-Li Lee1, Igor Rupniewski1, Christian M Beausejour1,2, Adam J Waite1, Nathaniel S Wang1, Kenneth A Kim1, Philip D Gregory1, Carl O Pabo1,2 & Edward J Rebar (2007);
An improved zinc-finger nuclease architecture for highly specific genome editing; "Nature Biotechnology" 25, 778 - 785