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Reference

Restriction enzyme reference

A searchable reference of common restriction enzymes: recognition site with the cut position marked, the overhang left behind, whether the enzyme is Type II or Type IIS, and which are everyday cloning workhorses. Filter by name or by recognition sequence.

Enzyme Recognition site Overhang Type Common
AatII GACGT^C 3′ overhang Type II
AflII C^TTAAG 5′ overhang Type II
AgeI A^CCGGT 5′ overhang Type II
AluI AG^CT blunt Type II yes
ApaI GGGCC^C 3′ overhang Type II
AscI GG^CGCGCC 5′ overhang Type II
AvrII C^CTAGG 5′ overhang Type II
BamHI G^GATCC 5′ overhang Type II yes
BbsI GAAGAC 5′ overhang Type IIS yes
BclI T^GATCA 5′ overhang Type II
BglII A^GATCT 5′ overhang Type II yes
BsaI GGTCTC 5′ overhang Type IIS yes
BsmBI CGTCTC 5′ overhang Type IIS yes
BspEI T^CCGGA 5′ overhang Type II
BspQI GCTCTTC 5′ overhang Type IIS
BsrGI T^GTACA 5′ overhang Type II
BstBI TT^CGAA 5′ overhang Type II
ClaI AT^CGAT 5′ overhang Type II
DdeI C^TNAG 5′ overhang Type II
DraI TTT^AAA blunt Type II
EagI C^GGCCG 5′ overhang Type II
EcoRI G^AATTC 5′ overhang Type II yes
EcoRV GAT^ATC blunt Type II yes
FseI GGCCGG^CC 3′ overhang Type II
HaeIII GG^CC blunt Type II yes
HhaI GCG^C 3′ overhang Type II
HindIII A^AGCTT 5′ overhang Type II yes
HinfI G^ANTC 5′ overhang Type II
HpaI GTT^AAC blunt Type II
HpaII C^CGG 5′ overhang Type II
KpnI GGTAC^C 3′ overhang Type II yes
MboI ^GATC 5′ overhang Type II
MfeI C^AATTG 5′ overhang Type II
MluI A^CGCGT 5′ overhang Type II
MspI C^CGG 5′ overhang Type II
NarI GG^CGCC 5′ overhang Type II
NcoI C^CATGG 5′ overhang Type II yes
NdeI CA^TATG 5′ overhang Type II yes
NheI G^CTAGC 5′ overhang Type II yes
NotI GC^GGCCGC 5′ overhang Type II yes
NruI TCG^CGA blunt Type II
NsiI ATGCA^T 3′ overhang Type II
PacI TTAAT^TAA 3′ overhang Type II
PciI A^CATGT 5′ overhang Type II
PmeI GTTT^AAAC blunt Type II
PstI CTGCA^G 3′ overhang Type II yes
PvuII CAG^CTG blunt Type II yes
RsaI GT^AC blunt Type II
SacI GAGCT^C 3′ overhang Type II yes
SacII CCGC^GG 3′ overhang Type II
SalI G^TCGAC 5′ overhang Type II yes
SapI GCTCTTC 5′ overhang Type IIS
Sau3AI ^GATC 5′ overhang Type II yes
SbfI CCTGCA^GG 3′ overhang Type II
ScaI AGT^ACT blunt Type II
SmaI CCC^GGG blunt Type II yes
SnaBI TAC^GTA blunt Type II
SpeI A^CTAGT 5′ overhang Type II yes
SphI GCATG^C 3′ overhang Type II
StuI AGG^CCT blunt Type II
SwaI ATTT^AAAT blunt Type II
TaqI T^CGA 5′ overhang Type II
XbaI T^CTAGA 5′ overhang Type II yes
XhoI C^TCGAG 5′ overhang Type II yes
XmaI C^CCGGG 5′ overhang Type II

65 enzymes listed

The caret marks the top-strand cut for Type II enzymes. Type IIS enzymes cut a fixed distance outside their recognition site, so no caret is shown; use the restriction sites tool to place their exact cut on a sequence.

A quick reference for cloning design. To find where these enzymes cut a specific sequence, or to scan for unique and non-cutting sites, use the restriction sites tool. Recognition sites are written 5′ to 3′ on the top strand.

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Frequently asked

How do I find where an enzyme cuts my sequence?

This page is a lookup table, not a scanner. Paste your sequence into the restriction sites tool to map cut positions, count fragments and find unique or non-cutting enzymes.

Why is there no cut mark on some enzymes?

Type IIS enzymes recognise one sequence but cut a defined distance away from it, so a caret inside the site would be misleading. The restriction sites tool places their cut correctly on a real sequence.

Do two enzymes with the same site cut identically?

Isoschizomers share a recognition sequence but may cut at different positions or leave different ends, and they differ in star activity and buffer needs. Check the supplier’s data for the exact enzyme you will use.

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