SfaN I

Enzyme nameSfaN I
PrototypeSfaNI
Known analogsLweI
SKUSE-E165
Turbo versionAvailable
High-concentration versionNot available
Restriction typeType 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'… GCATC(N)5▼ …3'
3'… CGTAG(N)9▲ …5'
SourceAn E.coli strain that carries the cloned SfaN I gene from Streptococcus faecalis N
Optimal bufferSE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
102510075025
Unit definitionOne 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 onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 300 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer O
References
  1. Y.E. Tomilova, M.A. Abdurashitov, L.N. Golikova, N.A. Netesova, S.Kh. Degtyarev Cloning of the Genes for DNA Methyltransferases of the SfaNI and Bst19I Restriction--Modification Systems and Primary Structure Analysis of Protein Products // Molecular Biology (Moscow), Vol. 38, No.6, 997-1004 (2004). (In Russian)
  2. G.D. Serov, S.Kh. Degtyarev, L.I. Puchkova, L.L. Korsun, N.I. Rechkunova Streptococcus faecalis - a strain producer of restriction endonuclease SfaNI. // Soviet Union patent SU 1645293 (1991). (In Russian)
  3. O.A. Zhilkina, V.E. Repin, S.Kh. Degtyarev Effect of some cultivation parameters on the yield of resctriction endonuclease SfaNI. // Applied Biochemistry and Microbiology (Moscow), Vol 27, No. 4, 486-490 (1991). (In Russian)
  4. O.A. Zhilkina, V.E. Repin, S.Kh. Degtyarev Preparation method of restriction endonuclease SfaNI, that recognizes and cleaves DNA sequence 5'-GCATC(N)5^, 3'-CGTAG(N)9^. // Soviet Union patent SU 1442546 (1988). (In Russian)
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