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| Enzyme | Recognition Site | Cut Count | Positions (bp) |
|---|---|---|---|
| AgsI | TTSAA | 2 cut(s) | 12, 61 |
| ApeKI | GCWGC | 1 cut(s) | 23 |
| AsuHPI | GGTGA | 1 cut(s) | 125 |
| BbvI | GCAGC | 1 cut(s) | 10 |
| BccI | CCATC | 1 cut(s) | 138 |
| BcgI | CGANNNNNNTGC | 1 cut(s) | 134 |
| BfaI | CTAG | 3 cut(s) | 18, 30, 92 |
| BglII | AGATCT | 1 cut(s) | 77 |
| BisI | GCNGC | 1 cut(s) | 24 |
| BlsI | GCNGC | 1 cut(s) | 25 |
| BmiI | GGNNCC | 1 cut(s) | 6 |
| BmsI | GCATC | 1 cut(s) | 125 |
| BseXI | GCAGC | 1 cut(s) | 10 |
| BsgI | GTGCAG | 1 cut(s) | 66 |
| Bsp143I | GATC | 1 cut(s) | 77 |
| BspLI | GGNNCC | 1 cut(s) | 6 |
| BssMI | GATC | 1 cut(s) | 77 |
| BstKTI | GATC | 1 cut(s) | 80 |
| BstMBI | GATC | 1 cut(s) | 77 |
| BstMWI | GCNNNNNNNGC | 2 cut(s) | 32, 80 |
| BstV1I | GCAGC | 1 cut(s) | 10 |
| BstX2I | RGATCY | 1 cut(s) | 77 |
| BstYI | RGATCY | 1 cut(s) | 77 |
| CviJI | RGCY | 3 cut(s) | 5, 23, 129 |
| CviKI_1 | RGCY | 3 cut(s) | 5, 23, 129 |
| DpnI | GATC | 1 cut(s) | 79 |
| DpnII | GATC | 1 cut(s) | 77 |
| EcoT22I | ATGCAT | 2 cut(s) | 58, 118 |
| FaiI | YATR | 1 cut(s) | 97 |
| Fnu4HI | GCNGC | 1 cut(s) | 24 |
| Fsp4HI | GCNGC | 1 cut(s) | 24 |
| FspBI | CTAG | 3 cut(s) | 18, 30, 92 |
| FspEI | CC | 9 cut(s) | 4, 5, 21, 22, 40, 87, 99, 143, 149 |
| GluI | GCNGC | 1 cut(s) | 24 |
| HphI | GGTGA | 1 cut(s) | 125 |
| HpyAV | CCTTC | 1 cut(s) | 18 |
| HpyCH4V | TGCA | 3 cut(s) | 56, 83, 116 |
| HpyF10VI | GCNNNNNNNGC | 2 cut(s) | 32, 80 |
| Kzo9I | GATC | 1 cut(s) | 77 |
| LmnI | GCTCC | 1 cut(s) | 10 |
| LpnPI | CCDG | 1 cut(s) | 87 |
| Lsp1109I | GCAGC | 1 cut(s) | 10 |
| LweI | GCATC | 1 cut(s) | 125 |
| MaeI | CTAG | 3 cut(s) | 18, 30, 92 |
| MaeIII | GTNAC | 1 cut(s) | 64 |
| MalI | GATC | 1 cut(s) | 79 |
| MboI | GATC | 1 cut(s) | 77 |
| MflI | RGATCY | 1 cut(s) | 77 |
| Mph1103I | ATGCAT | 2 cut(s) | 58, 118 |
| MseI | TTAA | 1 cut(s) | 157 |
| MwoI | GCNNNNNNNGC | 2 cut(s) | 32, 80 |
| NdeII | GATC | 1 cut(s) | 77 |
| NlaIV | GGNNCC | 1 cut(s) | 6 |
| NmuCI | GTSAC | 1 cut(s) | 64 |
| NsiI | ATGCAT | 2 cut(s) | 58, 118 |
| PkrI | GCNGC | 1 cut(s) | 25 |
| PspN4I | GGNNCC | 1 cut(s) | 6 |
| PsuI | RGATCY | 1 cut(s) | 77 |
| SaqAI | TTAA | 1 cut(s) | 157 |
| SatI | GCNGC | 1 cut(s) | 24 |
| Sau3AI | GATC | 1 cut(s) | 77 |
| SfaNI | GCATC | 1 cut(s) | 125 |
| SgeI | CNNG | 7 cut(s) | 30, 42, 48, 81, 87, 104, 114 |
| SspI | AATATT | 1 cut(s) | 46 |
| SspMI | CTAG | 3 cut(s) | 18, 30, 92 |
| TaqI | TCGA | 1 cut(s) | 119 |
| Tru1I | TTAA | 1 cut(s) | 157 |
| Tru9I | TTAA | 1 cut(s) | 157 |
| TseFI | GTSAC | 1 cut(s) | 64 |
| TseI | GCWGC | 1 cut(s) | 23 |
| Tsp45I | GTSAC | 1 cut(s) | 64 |
| XspI | CTAG | 3 cut(s) | 18, 30, 92 |
| Zsp2I | ATGCAT | 2 cut(s) | 58, 118 |