Fok I

Enzyme name Fok I
Prototype FokI
Known analogs BseGI, BstF5 I
SKU SE-E247
Turbo version Not available
High-concentration version Not available
Restriction type Type IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GGATG(N)9▼ …3'
3'… CCTAC(N)13▲ …5'
Source An E.coli strain that carries the cloned Fok I gene from Flavobacterium okeanokoites
Optimal buffer SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
50502525100100
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA
Storage conditions 10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligation After 2-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer Y
Notes Overdigestions of > 5 units of Fok I per 1μg of DNA and incubations > 2 hours are not recommended.
References
  3. M.A. Abdurashitov, E.V. Kileva, N.M. Shinkarenko, A.V. Shevchenko, V.S. Dedkov, S.Kh.Degtyarev BstF5 I, an unusual isoschizomer of Fok I. // Gene, 172, 4951 (1996).
  4. S.K. Degtyarev, P.A. Belavin, V.E. Repin, E.G. Malyvin Preparation method of restriction endonuclease Fok I from Flavobacterium okeanokoites // Soviet Union patent SU1436160 (1988). (In Russian)
  5. External publication Sugisaki, H., Kanazawa, S. Gene 16: 73-78 (1981).
// Код для некролога