Table 3.
REase | Recognition site | Number of sites in pHia5ET | Sensitivity to m6A | Cleavage |
---|---|---|---|---|
BglII | AGaTCT | 1 | S | N |
Bsh1285I | CGRYCG R = A/G | 5 | S | N |
Bsu15I | ATCGAT | 2 | S | N |
CviQI | GTAC | 3 | S | N |
DraI | TTTAAA | 2 | S | N |
Eco32I | GATATC | 0 (1)b | S | N |
EcoRI | GAATTC | 0 (1)b | S | N |
HincII | GTYRAC | 1 (GTTAAC) | S | N |
HindIII | AaGCTT | 0 (1)b | S | N |
HinfI | GANTC | 19 | S | N |
HphI | GGTGA/TCACC | 16 | S | N |
KspAI | GTTAAC | 1 | S | N |
MboIa | GATC | 24 | S | N |
MluI | db ACGCGT | 1 | S | N |
MlyI | GAGTC/GACTC | 9 | S | N |
MunI | CAATTG | 1 | S | N |
NlaIII | CATG | 24 | S | N |
PaeI | GCATGC | 1 | S | N |
PstI | CTGCAG | 0 (2)b | S | N |
PvuII | dbCAGCTG | 3 | S | N |
SspI | aATATT | 3 | S | N |
VspI | ATTAAT | 5 | S | N |
XbaI | TCTaGA | 1 | S | N |
XhoI | CTCGAG | 1 | S | N |
TaiI | ACGT | 16 | S | N |
TaqI | TCGA | 16 | S | N |
TfiI | GAWTC | 10 | S | N |
AluI | AGCT | 24 | S | P, L |
TasI | AATT | 28 | S | P, L |
Tru1I | TTAA | 29 | S | P, L |
CaiI | CAGNNNCTG | 1 | U | N |
CseI | GACGC | 12 | U | N |
Eam1104I | GAAGAG | 3 | U | N |
Eco147I | AGGCCT | 1 | U | N |
Eco47III | AGCGCT | 3 | U | N |
Eco91I | GGTNACC | 1 | U | N |
LweI | GCATC | 24 | U | N |
Mph1103I | ATGCAT | 3 | U | N |
NdeI | CaTATG | 1 | U | N |
PfeI | GAWTC | 10 | U | N |
PscI | ACATGT | 1 | U | N |
SchI | GAGTC/GACTC | 9 | U | N |
XmiI | GTMKAC | 1 | U | N |
XapI | RAATTY | 6 | U | N |
AdeI | CACNNNGTG | 1 | U | N |
BpiI | GAAGAC | 5 | U | N |
Bsp119I | TTCGAA | 1 | U | N |
BcnI | CCSGG | 12 | (−) | Y |
BglI | GGCN5GCC | 1 | (−) | Y |
Bme1390I | CCNGG | 21 | (−) | Y |
Bpu1102I | GCTNAGC | 2 | U | Y |
Bsh1236I | CGCG | 37 | (−) | Y |
Bsp143Ia | GATC | 24 | Not | Y |
Bsp120I | GGGCCC | 1 | (−) | Y |
Bsp68I | TCGCGA | 1 | Not | Y |
BspLI | GGNNCC | 20 | (−) | Y |
BspOI | GCTAGC | 0 (2)b | U | Y |
Bst1107I | GTATAC | 1 | U | Y |
BsuRI | GGCC | 22 | (−) | Y |
Csp6I | GTAC | 3 | U | Y |
DpnIa | Gm6ATC | 24 | Not | Y |
Eco130I | CCWWGG | 1 | (−) | Y |
Eco88I | CYCGRG | 2 | (−) | Y |
EcoO109I | RGGNCCY | 3 | (−) | Y |
FspBI | CTAG | 6 | U | Y |
Hin1I | GRCGYC | 5 | (−) | Y |
Hin6I | GCGC | 32 | (−) | Y |
HpyF10VI | GCN7GC | 41 | (−) | Y |
MbiI | GAGCGG/CCGCTC | 4 | U | Y |
MspI | CCGG | 29 | (−) | Y |
NheI | GCTAGC | 0 (2)b | U | Y |
PvuI | CGATCG | 1 | Not | Y |
SmaI | CCCGGG | 1 | (−) | Y |
aTested in λ DNA dam- in vitro methylated with the Hia5 enzyme. bnumber of recognition sites in plasmid pAltOKI.
Y: complete cleavage; N: no digestion; P: partial digestion, L: plasmid linearized; S: sensitive to m6A; U: unknown sensitivity to m6A; Not: not sensitive to m6A; (−): absence of adenine residues in recognition sequence; db: methylation of both strands required to block cleavage; A: the enzyme will not cut if the marked adenine is methylated; a: the enzyme is not sensitive to methylation of the marked adenine; A: there are no data on the sensitivity of the enzyme to methylation of the unmarked adenine.