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| Enzyme | Recognition Site | Cut Count | Positions (bp) |
|---|---|---|---|
| AciI | CCGC | 2 cut(s) | 51, 76 |
| AcoI | YGGCCR | 1 cut(s) | 64 |
| AfiI | CCNNNNNNNGG | 1 cut(s) | 135 |
| AlwNI | CAGNNNCTG | 1 cut(s) | 25 |
| AoxI | GGCC | 1 cut(s) | 64 |
| BanII | GRGCYC | 1 cut(s) | 141 |
| BauI | CACGAG | 1 cut(s) | 140 |
| BceAI | ACGGC | 1 cut(s) | 18 |
| BmrI | ACTGGG | 1 cut(s) | 13 |
| BmuI | ACTGGG | 1 cut(s) | 13 |
| BsaJI | CCNNGG | 1 cut(s) | 61 |
| Bsc4I | CCNNNNNNNGG | 1 cut(s) | 135 |
| Bse1I | ACTGG | 1 cut(s) | 19 |
| BseDI | CCNNGG | 1 cut(s) | 61 |
| BseGI | GGATG | 1 cut(s) | 14 |
| BseLI | CCNNNNNNNGG | 1 cut(s) | 135 |
| BseNI | ACTGG | 1 cut(s) | 19 |
| BshFI | GGCC | 1 cut(s) | 66 |
| BslFI | GGGAC | 2 cut(s) | 7, 112 |
| BslI | CCNNNNNNNGG | 1 cut(s) | 135 |
| BsmFI | GGGAC | 2 cut(s) | 7, 112 |
| BsnI | GGCC | 1 cut(s) | 66 |
| Bsp1286I | GDGCHC | 1 cut(s) | 141 |
| BspACI | CCGC | 2 cut(s) | 51, 76 |
| BspANI | GGCC | 1 cut(s) | 66 |
| BsrI | ACTGG | 1 cut(s) | 19 |
| BssECI | CCNNGG | 1 cut(s) | 61 |
| BssSI | CACGAG | 1 cut(s) | 140 |
| Bst2BI | CACGAG | 1 cut(s) | 140 |
| BstF5I | GGATG | 1 cut(s) | 14 |
| BsuRI | GGCC | 1 cut(s) | 66 |
| BtsCI | GGATG | 1 cut(s) | 14 |
| CaiI | CAGNNNCTG | 1 cut(s) | 25 |
| CviAII | CATG | 2 cut(s) | 40, 68 |
| CviJI | RGCY | 4 cut(s) | 60, 66, 119, 139 |
| CviKI_1 | RGCY | 4 cut(s) | 60, 66, 119, 139 |
| EaeI | YGGCCR | 1 cut(s) | 64 |
| Eco24I | GRGCYC | 1 cut(s) | 141 |
| EcoT38I | GRGCYC | 1 cut(s) | 141 |
| FaeI | CATG | 2 cut(s) | 43, 71 |
| FaiI | YATR | 3 cut(s) | 41, 69, 148 |
| FaqI | GGGAC | 2 cut(s) | 7, 112 |
| FatI | CATG | 2 cut(s) | 39, 67 |
| FauI | CCCGC | 1 cut(s) | 83 |
| FriOI | GRGCYC | 1 cut(s) | 141 |
| HaeIII | GGCC | 1 cut(s) | 66 |
| Hin1II | CATG | 2 cut(s) | 43, 71 |
| HinfI | GANTC | 2 cut(s) | 110, 125 |
| Hpy188I | TCNGA | 2 cut(s) | 98, 115 |
| HpyCH4V | TGCA | 1 cut(s) | 4 |
| Hsp92II | CATG | 2 cut(s) | 43, 71 |
| LmnI | GCTCC | 1 cut(s) | 76 |
| LpnPI | CCDG | 3 cut(s) | 32, 38, 92 |
| MaeIII | GTNAC | 1 cut(s) | 10 |
| MhlI | GDGCHC | 1 cut(s) | 141 |
| MluCI | AATT | 1 cut(s) | 6 |
| MlyI | GAGTC | 1 cut(s) | 134 |
| MnlI | CCTC | 1 cut(s) | 71 |
| MseI | TTAA | 1 cut(s) | 90 |
| NlaIII | CATG | 2 cut(s) | 43, 71 |
| NmeAIII | GCCGAG | 1 cut(s) | 42 |
| NmuCI | GTSAC | 1 cut(s) | 10 |
| PfeI | GAWTC | 1 cut(s) | 110 |
| PleI | GAGTC | 1 cut(s) | 133 |
| PpsI | GAGTC | 1 cut(s) | 133 |
| PstNI | CAGNNNCTG | 1 cut(s) | 25 |
| SaqAI | TTAA | 1 cut(s) | 90 |
| SchI | GAGTC | 1 cut(s) | 134 |
| SduI | GDGCHC | 1 cut(s) | 141 |
| SgeI | CNNG | 8 cut(s) | 31, 37, 52, 74, 80, 87, 91, 97 |
| Sse9I | AATT | 1 cut(s) | 6 |
| SsiI | CCGC | 2 cut(s) | 51, 76 |
| TasI | AATT | 1 cut(s) | 6 |
| TfiI | GAWTC | 1 cut(s) | 110 |
| Tru1I | TTAA | 1 cut(s) | 90 |
| Tru9I | TTAA | 1 cut(s) | 90 |
| TseFI | GTSAC | 1 cut(s) | 10 |
| Tsp45I | GTSAC | 1 cut(s) | 10 |