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| Enzyme | Recognition Site | Cut Count | Positions (bp) |
|---|---|---|---|
| AgsI | TTSAA | 4 cut(s) | 32, 154, 163, 195 |
| AsuHPI | GGTGA | 2 cut(s) | 191, 235 |
| BarI | GAAGNNNNNNTAC | 2 cut(s) | 128, 160 |
| BmiI | GGNNCC | 1 cut(s) | 126 |
| BsaAI | YACGTR | 1 cut(s) | 248 |
| BsaJI | CCNNGG | 1 cut(s) | 10 |
| BsaXI | ACNNNNNCTCC | 2 cut(s) | 6, 36 |
| Bse3DI | GCAATG | 1 cut(s) | 225 |
| BseDI | CCNNGG | 1 cut(s) | 10 |
| BseMI | GCAATG | 1 cut(s) | 225 |
| BseMII | CTCAG | 1 cut(s) | 101 |
| BspCNI | CTCAG | 1 cut(s) | 102 |
| BspLI | GGNNCC | 1 cut(s) | 126 |
| BsrDI | GCAATG | 1 cut(s) | 225 |
| BssECI | CCNNGG | 1 cut(s) | 10 |
| BssT1I | CCWWGG | 1 cut(s) | 10 |
| BstBAI | YACGTR | 1 cut(s) | 248 |
| BstC8I | GCNNGC | 1 cut(s) | 200 |
| BstDEI | CTNAG | 3 cut(s) | 57, 89, 110 |
| BstEII | GGTNACC | 1 cut(s) | 179 |
| BstPI | GGTNACC | 1 cut(s) | 179 |
| Cac8I | GCNNGC | 1 cut(s) | 200 |
| CviAII | CATG | 1 cut(s) | 64 |
| CviJI | RGCY | 2 cut(s) | 242, 254 |
| CviKI_1 | RGCY | 2 cut(s) | 242, 254 |
| DdeI | CTNAG | 3 cut(s) | 57, 89, 110 |
| Eco130I | CCWWGG | 1 cut(s) | 10 |
| Eco91I | GGTNACC | 1 cut(s) | 179 |
| EcoO65I | GGTNACC | 1 cut(s) | 179 |
| EcoT14I | CCWWGG | 1 cut(s) | 10 |
| ErhI | CCWWGG | 1 cut(s) | 10 |
| FaeI | CATG | 1 cut(s) | 67 |
| FaiI | YATR | 5 cut(s) | 45, 65, 75, 190, 208 |
| FatI | CATG | 1 cut(s) | 63 |
| Hin1II | CATG | 1 cut(s) | 67 |
| HphI | GGTGA | 2 cut(s) | 191, 235 |
| Hpy188I | TCNGA | 2 cut(s) | 104, 111 |
| HpyAV | CCTTC | 2 cut(s) | 115, 208 |
| HpyCH4IV | ACGT | 1 cut(s) | 247 |
| HpyF3I | CTNAG | 3 cut(s) | 57, 89, 110 |
| HpySE526I | ACGT | 1 cut(s) | 247 |
| Hsp92II | CATG | 1 cut(s) | 67 |
| LpnPI | CCDG | 1 cut(s) | 184 |
| MaeII | ACGT | 1 cut(s) | 247 |
| MaeIII | GTNAC | 3 cut(s) | 22, 47, 179 |
| MboII | GAAGA | 4 cut(s) | 107, 148, 166, 179 |
| MnlI | CCTC | 2 cut(s) | 105, 219 |
| NlaIII | CATG | 1 cut(s) | 67 |
| NlaIV | GGNNCC | 1 cut(s) | 126 |
| NmuCI | GTSAC | 2 cut(s) | 47, 179 |
| Ppu21I | YACGTR | 1 cut(s) | 248 |
| PspEI | GGTNACC | 1 cut(s) | 179 |
| PspN4I | GGNNCC | 1 cut(s) | 126 |
| SetI | ASST | 4 cut(s) | 130, 181, 230, 250 |
| SgeI | CNNG | 9 cut(s) | 23, 44, 76, 94, 196, 207, 211, 223, 258 |
| StyI | CCWWGG | 1 cut(s) | 10 |
| TaiI | ACGT | 1 cut(s) | 250 |
| TseFI | GTSAC | 2 cut(s) | 47, 179 |
| Tsp45I | GTSAC | 2 cut(s) | 47, 179 |
| TspDTI | ATGAA | 1 cut(s) | 180 |