| Enzyme | Cleavage Site | Overhang | Iso-/Neoschizomer at JBS | Other Iso-/Neoschizomers |
| Iso | Neo (Cleavage Site) | Iso | Neo |
| BamH I | 5 ' - G | G A T C C - 3 ' 3 ' - C C T A G | G - 5 ' | 5'-GATC |
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Ban III
| 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Bsa29 I, Bsp106 I, BspD I, BspX I, Bsu15 I, BsuTU I, Cla I, Zho I |
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| Bbu I | 5 ' - G C A T G | C - 3 ' 3 ' - C | G T A C G - 5 ' | CATG-3' | Sph I |
| Pae I, SpaH I |
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| Bcl I | 5 ' - T | G A T C A - 3 ' 3 ' - A C T A G | T - 5 ' | 5'-GATC |
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| Fba I, Ksp22 I |
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| Bfr I | 5 ' - C | T T A A G - 3 ' 3 ' - G A A T T | C - 5 ' | 5'-TTAA | MspC I |
| Afl II, BspT I, Bst98 I, Vha464 I |
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| BfuC I | 5 ' - | G A T C - 3 ' 3 ' - C T A G | - 5 ' | 5'-GATC | Mbo I
Sau3A I | Dpn I
| Bsp143 I, BstEN II, BstMB I, Dpn II, Kzo9 I, Nde II | BstKT I |
| Bgl I | 5 ' - G C C N N N N | N G G C - 3 ' 3 ' - C G G N | N N N N C C G - 5 ' | NNN-3' |
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| Bgl II | 5 ' - A | G A T C T - 3 ' 3 ' - T C T A G | A - 5 ' | 5'-GATC |
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| Bmt I | 5 ' - G C T A G | C - 3 ' 3 ' - C | G A T C G - 5 ' | CTAG-3' |
| Nhe I
5 ' - G | C T A G C - 3 ' 3 ' - C G A T C | G - 5 ' |
| AsuNH I |
| Bpu14 I | 5 ' - T T | C G A A - 3 ' 3 ' - A A G C | T T - 5 ' | 5'-CG | Asu II |
| Bsp119 I, BspT104 I, BstB I, Csp45 I, Nsp V, Sfu I |
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| Bsa29 I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Ban III, Bsp106 I, BspD I, BspX I, Bsu15 I, BsuTU I, Cla I, Zho I |
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| Bse118 I | 5 ' - R C | C G G Y - 3 ' 3 ' - Y G G C | C R - 5 ' | 5'-CG | BssA I |
| BsrF I, Cfr10 I |
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| BseA I | 5 ' - T | C C G G A - 3 ' 3 ' - A G G C C | T - 5 ' | 5'-CCGG |
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| Acc III, Bsp13 I, BspE I, Kpn2 I, Mro I |
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| BseB I | 5 ' - C C | W G G - 3 ' 3 ' - G G W | C C - 5 ' | 5'-W |
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| BstN I, BstO I, Bst2U I, Mva I | Ajn I, EcoR II, Psp6 I, PspG I |
| BseC I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG |
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| Ban III, Bsa29 I, Bsp106 I, BspD I, BspX I, Bsu15 I, BsuTU I, Cla I, Zho I |
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| BshF I | | blunt |
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| BspAN I, BsuR I, Hae III, Pal I, Pho I |
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| BshT I | 5 ' - A | C C G G T - 3 ' 3 ' - T G G C C | A - 5 ' | 5'-CCGG | CspA I |
| Age I, AsiA I, PinA I |
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| BsiHKC I | 5 ' - C | Y C G R G - 3 ' 3 ' - G R G C Y | C - 5 ' | 5'-YCGR | BspLU4 I |
| Ama87 I, Ava I, BsoB I, Eco88 I, Nsp III |
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| BsiS I | 5 ' - C | C G G - 3 ' 3 ' - G G C | C - 5 ' | 5'-CG |
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| Hap II, Hpa II, Msp I |
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| BsoB I | 5 ' - C | Y C G R G - 3 ' 3 ' - G R G C Y | C - 5 ' | 5'-YCGR | BspLU4 I |
| Ama87 I, Ava I, BsiHKC I, Eco88 I, Nsp III |
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| Bsp106 I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Ban III, Bsa29 I, BspD I, BspX I, Bsu15 I, BsuTU I, Cla I, Zho I |
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| Bsp119 I | 5 ' - T T | C G A A - 3 ' 3 ' - A A G C | T T - 5 ' | 5'-CG | Asu II |
| Bpu14 I, BspT104 I, BstB I, Csp45 I, Nsp V, Sfu I |
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| Bsp13 I | 5 ' - T | C C G G A - 3 ' 3 ' - A G G C C | T - 5 ' | 5'-CCGG | BseA I |
| Acc III, BspE I, Kpn2 I, Mro I |
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| Bsp143 I | 5 ' - | G A T C - 3 ' 3 ' - C T A G | - 5 ' | 5'-GATC | Mbo I
Sau3A I | Dpn I
| BfuC I, BstEN II, BstMB I, Dpn II, Kzo9 I, Nde II | BstKT I |
| Bsp19 I | 5 ' - C | C A T G G - 3 ' 3 ' - G G T A C | C - 5 ' | 5'-CATG | Nco I |
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| Bsp68 I | 5 ' - T C G | C G A - 3 ' 3 ' - A G C | G C T - 5 ' | blunt | Nru I |
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| BspAN I | | blunt | BshF I |
| BsuR I, Hae III, Pal I, Pho I |
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| BspD I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Ban III, Bsa29 I, Bsp106 I, BspX I, Bsu15 I, BsuTU I, Cla I, Zho I |
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| BspE I | 5 ' - T | C C G G A - 3 ' 3 ' - A G G C C | T - 5 ' | 5'-CCGG | BseA I |
| Acc III, Bsp13 I, Kpn2 I, Mro I |
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| BspLU11 I | 5 ' - A | C A T G T - 3 ' 3 ' - T G T A C | A - 5 ' | 5'-CATG |
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| Pci I, Psc I |
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| BspT I | 5 ' - C | T T A A G - 3 ' 3 ' - G A A T T | C - 5 ' | 5'-TTAA | MspC I |
| Afl II, Bfr I, Bst98 I, Vha464 I |
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| BspT104 I | 5 ' - T T | C G A A - 3 ' 3 ' - A A G C | T T - 5 ' | 5'-CG | Asu II |
| Bpu14 I, Bsp119 I, BstB I, Csp45 I, Nsp V, Sfu I |
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| BspX I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Ban III, Bsa29 I, Bsp106 I, BspD I, Bsu15 I, BsuTU I, Cla I, Zho I |
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| BsrF I | 5 ' - R C | C G G Y - 3 ' 3 ' - Y G G C | C R - 5 ' | 5'-CG | Bss A I |
| Bse118 I, Cfr10 I |
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| BssA I | 5 ' - R C | C G G Y - 3 ' 3 ' - Y G G C | C R - 5 ' | 5'-CG |
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| Bse118 I, BsrF I, Cfr10 I |
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| BssH I | 5 ' - C | T C G A G - 3 ' 3 ' - G A G C T | C - 5 ' | 5'-TCGA | Sla I |
| PaeR7 I, Sfr274 I, Tli I, Xho I |
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| BssT1 I | 5 ' - C | C W W G G - 3 ' 3 ' - G G W W C | C - 5 ' | 5'-CWWG | Sty I |
| Eco130 I, EcoT14 I, Erh I |
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| Bst2U I | 5 ' - C C | W G G - 3 ' 3 ' - G G W | C C - 5 ' | 5'-W | Bse B I |
| BstN I, BstO I, Mva I | Ajn I, EcoR II, Psp6 I, PspG I |
| Bst98 I | 5 ' - C | T T A A G - 3 ' 3 ' - G A A T T | C - 5 ' | 5'-TTAA | MspC I |
| Afl II, Bfr I, BspT I, Vha464 I |
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| BstB I | 5 ' - T T | C G A A - 3 ' 3 ' - A A G C | T T - 5 ' | 5'-CG | Asu II |
| Bpu14 I, Bsp119 I, BspT104 I, Csp45 I, Nsp V ,Sfu I |
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| BstE II | 5 ' - G | G T N A C C - 3 ' 3 ' - C C A N T G | G - 5 ' | 5'-GTNAC |
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| BstP I, Eco91 I, EcoO65 I, PspE I |
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| BstEN II | 5 ' - | G A T C - 3 ' 3 ' - C T A G | - 5 ' | 5'-GATC | Mbo I
Sau3A I | Dpn I
| BfuC I, Bsp143 I, BstMB I, Dpn II, Kzo9 I, Nde II | BstKT I |
| BstKT I | 5 ' - G A T | C - 3 ' 3 ' - C | T A G - 5 ' | AT-3' |
| Mbo I Sau3A I
5 ' - | G A T C - 3 ' 3 ' - C T A G | - 5 ' |
| BfuC I, Bsp143 I, BstEN II, BstMB I, Dpn II, Kzo9 I, Nde II |
Dpn I
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| BstMA I | 5 ' - C T G C A | G - 3 ' 3 ' - G | A C G T C - 5 ' | TGCA-3' | Pst I |
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| BstMB I | 5 ' - | G A T C - 3 ' 3 ' - C T A G | - 5 ' | 5'-GATC | Mbo I
Sau3A I | Dpn I
| BfuC I, Bsp143 I, BstEN II, Dpn II, Kzo9 I, Nde II | BstKT I |
| BstN I | 5 ' - C C | W G G - 3 ' 3 ' - G G W | C C - 5 ' | 5'-W | BseB I |
| BstO I, Bst2U I, Mva I | Ajn I, EcoR II, Psp6 I, PspG I |
| BstO I | 5 ' - C C | W G G - 3 ' 3 ' - G G W | C C - 5 ' | 5'-W | BseB I |
| BstN I, Bst2U I, Mva I | Ajn I, EcoR II, Psp6 I, PspG I |
| BstP I | 5 ' - G | G T N A C C - 3 ' 3 ' - C C A N T G | G - 5 ' | 5'-GTNAC | BstE II |
| Eco91 I, EcoO65 I, PspE I |
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| BstSN I | 5 ' - T A C | G T A - 3 ' 3 ' - A T G | C A T - 5 ' | blunt | SnaB I |
| Eco105 I |
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| Bsu15 I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Ban III, Bsa29 I, Bsp106 I, BspD I, BspX I, BsuTU I, Cla I, Zho I |
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| BsuR I | | blunt | BshF I |
| BspAN I, Hae III, Pal I, Pho I |
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| BsuTU I | 5 ' - A T | C G A T - 3 ' 3 ' - T A G C | T A - 5 ' | 5'-CG | BseC I |
| Ban III, Bsa29 I, Bsp106 I, BspD I, BspX I, Bsu15 I, Cla I, Zho I |
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