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| Enzyme | Recognition Site | Cut Count | Positions (bp) |
|---|---|---|---|
| AciI | CCGC | 3 cut(s) | 17, 21, 365 |
| AcoI | YGGCCR | 1 cut(s) | 18 |
| AcuI | CTGAAG | 2 cut(s) | 87, 321 |
| AfiI | CCNNNNNNNGG | 1 cut(s) | 195 |
| AgsI | TTSAA | 2 cut(s) | 161, 341 |
| AluBI | AGCT | 3 cut(s) | 174, 184, 329 |
| AluI | AGCT | 3 cut(s) | 174, 184, 329 |
| Alw26I | GTCTC | 2 cut(s) | 134, 282 |
| AoxI | GGCC | 2 cut(s) | 18, 246 |
| AspS9I | GGNCC | 1 cut(s) | 247 |
| AsuC2I | CCSGG | 1 cut(s) | 250 |
| AsuHPI | GGTGA | 1 cut(s) | 161 |
| BccI | CCATC | 3 cut(s) | 152, 198, 263 |
| BclI | TGATCA | 1 cut(s) | 151 |
| BcnI | CCSGG | 1 cut(s) | 250 |
| BcoDI | GTCTC | 2 cut(s) | 134, 282 |
| BisI | GCNGC | 3 cut(s) | 18, 21, 366 |
| BlsI | GCNGC | 3 cut(s) | 19, 22, 367 |
| Bme1390I | CCNGG | 1 cut(s) | 250 |
| BmgT120I | GGNCC | 1 cut(s) | 247 |
| BmrFI | CCNGG | 1 cut(s) | 250 |
| BmsI | GCATC | 1 cut(s) | 310 |
| Bpu10I | CCTNAGC | 2 cut(s) | 170, 330 |
| BpuEI | CTTGAG | 1 cut(s) | 158 |
| BpuMI | CCSGG | 1 cut(s) | 250 |
| BsaI | GGTCTC | 1 cut(s) | 134 |
| Bsc4I | CCNNNNNNNGG | 1 cut(s) | 195 |
| BseLI | CCNNNNNNNGG | 1 cut(s) | 195 |
| BseX3I | CGGCCG | 1 cut(s) | 18 |
| Bsh1285I | CGRYCG | 1 cut(s) | 21 |
| BshFI | GGCC | 2 cut(s) | 20, 248 |
| BsiEI | CGRYCG | 1 cut(s) | 21 |
| BsiSI | CCGG | 1 cut(s) | 250 |
| BslI | CCNNNNNNNGG | 1 cut(s) | 195 |
| BsmAI | GTCTC | 2 cut(s) | 134, 282 |
| BsnI | GGCC | 2 cut(s) | 20, 248 |
| Bso31I | GGTCTC | 1 cut(s) | 134 |
| Bsp143I | GATC | 1 cut(s) | 151 |
| BspACI | CCGC | 3 cut(s) | 17, 21, 365 |
| BspANI | GGCC | 2 cut(s) | 20, 248 |
| BspQI | GCTCTTC | 1 cut(s) | 297 |
| BspTNI | GGTCTC | 1 cut(s) | 134 |
| BssMI | GATC | 1 cut(s) | 151 |
| Bst4CI | ACNGT | 1 cut(s) | 198 |
| Bst6I | CTCTTC | 2 cut(s) | 237, 297 |
| BstC8I | GCNNGC | 4 cut(s) | 25, 29, 255, 309 |
| BstDEI | CTNAG | 2 cut(s) | 170, 330 |
| BstKTI | GATC | 1 cut(s) | 154 |
| BstMAI | GTCTC | 2 cut(s) | 134, 282 |
| BstMBI | GATC | 1 cut(s) | 151 |
| BstMCI | CGRYCG | 1 cut(s) | 21 |
| BstMWI | GCNNNNNNNGC | 1 cut(s) | 254 |
| BstSCI | CCNGG | 1 cut(s) | 248 |
| BstZI | CGGCCG | 1 cut(s) | 18 |
| BsuRI | GGCC | 2 cut(s) | 20, 248 |
| Cac8I | GCNNGC | 4 cut(s) | 25, 29, 255, 309 |
| CciNI | GCGGCCGC | 1 cut(s) | 18 |
| Cfr13I | GGNCC | 1 cut(s) | 247 |
| CseI | GACGC | 1 cut(s) | 66 |
| DdeI | CTNAG | 2 cut(s) | 170, 330 |
| DpnI | GATC | 1 cut(s) | 153 |
| DpnII | GATC | 1 cut(s) | 151 |
| EaeI | YGGCCR | 1 cut(s) | 18 |
| EagI | CGGCCG | 1 cut(s) | 18 |
| Eam1104I | CTCTTC | 2 cut(s) | 237, 297 |
| EarI | CTCTTC | 2 cut(s) | 237, 297 |
| EclXI | CGGCCG | 1 cut(s) | 18 |
| Eco31I | GGTCTC | 1 cut(s) | 134 |
| Eco52I | CGGCCG | 1 cut(s) | 18 |
| Eco57I | CTGAAG | 2 cut(s) | 87, 321 |
| FaiI | YATR | 6 cut(s) | 92, 111, 131, 228, 233, 371 |
| FalI | AAGNNNNNCTT | 2 cut(s) | 275, 307 |
| FbaI | TGATCA | 1 cut(s) | 151 |
| Fnu4HI | GCNGC | 3 cut(s) | 18, 21, 366 |
| Fsp4HI | GCNGC | 3 cut(s) | 18, 21, 366 |
| GluI | GCNGC | 3 cut(s) | 18, 21, 366 |
| HaeIII | GGCC | 2 cut(s) | 20, 248 |
| HapII | CCGG | 1 cut(s) | 250 |
| HgaI | GACGC | 1 cut(s) | 66 |
| HinfI | GANTC | 1 cut(s) | 116 |
| HpaII | CCGG | 1 cut(s) | 250 |
| HphI | GGTGA | 1 cut(s) | 161 |
| Hpy166II | GTNNAC | 1 cut(s) | 201 |
| Hpy188I | TCNGA | 2 cut(s) | 40, 56 |
| Hpy188III | TCNNGA | 1 cut(s) | 120 |
| Hpy8I | GTNNAC | 1 cut(s) | 201 |
| HpyAV | CCTTC | 3 cut(s) | 35, 200, 343 |
| HpyCH4III | ACNGT | 1 cut(s) | 198 |
| HpyCH4V | TGCA | 2 cut(s) | 4, 358 |
| HpyF10VI | GCNNNNNNNGC | 1 cut(s) | 254 |
| HpyF3I | CTNAG | 2 cut(s) | 170, 330 |
| Ksp22I | TGATCA | 1 cut(s) | 151 |
| Kzo9I | GATC | 1 cut(s) | 151 |
| LguI | GCTCTTC | 1 cut(s) | 297 |
| LpnPI | CCDG | 5 cut(s) | 32, 41, 105, 252, 263 |
| LweI | GCATC | 1 cut(s) | 310 |
| MalI | GATC | 1 cut(s) | 153 |
| MboI | GATC | 1 cut(s) | 151 |
| MboII | GAAGA | 2 cut(s) | 254, 314 |
| MnlI | CCTC | 3 cut(s) | 94, 203, 238 |
| MseI | TTAA | 1 cut(s) | 216 |
| MspI | CCGG | 1 cut(s) | 250 |
| MspR9I | CCNGG | 1 cut(s) | 250 |
| MwoI | GCNNNNNNNGC | 1 cut(s) | 254 |
| NciI | CCSGG | 1 cut(s) | 250 |
| NdeII | GATC | 1 cut(s) | 151 |
| NotI | GCGGCCGC | 1 cut(s) | 18 |
| PciSI | GCTCTTC | 1 cut(s) | 297 |
| PfeI | GAWTC | 1 cut(s) | 116 |
| PkrI | GCNGC | 3 cut(s) | 19, 22, 367 |
| PspPI | GGNCC | 1 cut(s) | 247 |
| SapI | GCTCTTC | 1 cut(s) | 297 |
| SaqAI | TTAA | 1 cut(s) | 216 |
| SatI | GCNGC | 3 cut(s) | 18, 21, 366 |
| Sau3AI | GATC | 1 cut(s) | 151 |
| Sau96I | GGNCC | 1 cut(s) | 247 |
| ScrFI | CCNGG | 1 cut(s) | 250 |
| SetI | ASST | 5 cut(s) | 130, 176, 186, 227, 331 |
| SfaNI | GCATC | 1 cut(s) | 310 |
| SmlI | CTYRAG | 1 cut(s) | 137 |
| SmoI | CTYRAG | 1 cut(s) | 137 |
| SsiI | CCGC | 3 cut(s) | 17, 21, 365 |
| StyD4I | CCNGG | 1 cut(s) | 248 |
| TaaI | ACNGT | 1 cut(s) | 198 |
| TauI | GCSGC | 3 cut(s) | 20, 23, 368 |
| TfiI | GAWTC | 1 cut(s) | 116 |
| Tru1I | TTAA | 1 cut(s) | 216 |
| Tru9I | TTAA | 1 cut(s) | 216 |
| TspDTI | ATGAA | 1 cut(s) | 98 |