* Blocked by Dam methylation.BclI is typically used at 50-55°C, but is 50% active at 37°C.
BsaBI (4081) 1 site
GATNNNNATCCTANNNNTAG
* Blocked by Dam methylation.
MfeI (3989) 1 site
CAATTGGTTAAC
HpaI (3980) 1 site
GTTAACCAATTG
BspDI (3844) 1 site
ATCGATTAGCTA
* Blocked by Dam methylation.
ClaI (3844) 1 site
ATCGATTAGCTA
* Blocked by Dam methylation.
NaeI (3592) 1 site
GCCGGCCGGCCG
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV (3590) 1 site
GCCGGCCGGCCG
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
DraIII (3489) 1 site
CACNNNGTGGTGNNNCAC
Sticky ends from different DraIII sites may not be compatible.
PflMI (3177) 1 site
CCANNNNNTGGGGTNNNNNACC
* Blocked by Dcm methylation.Sticky ends from different PflMI sites may not be compatible.
XmnI (2789) 1 site
GAANNNNTTCCTTNNNNAAG
ScaI (2670) 1 site
AGTACTTCATGA
BsaI (2251) 1 site
GGTCTCNCCAGAGN(N)4
Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°C.
MluI (61) 1 site
ACGCGTTGCGCA
HindIII (67) 1 site
AAGCTTTTCGAA
PspOMI (73) 1 site
GGGCCCCCCGGG
ApaI (77) 1 site
GGGCCCCCCGGG
ApaI can be used between 25°C and 37°C.
AbsI (79) 1 site
CCTCGAGGGGAGCTCC
PaeR7I (79) 1 site
CTCGAGGAGCTC
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI (79) 1 site
VCTCGAGBBGAGCTCV
XhoI (79) 1 site
CTCGAGGAGCTC
XbaI (91) 1 site
TCTAGAAGATCT
EagI (98) 1 site
CGGCCGGCCGGC
NotI (98) 1 site
GCGGCCGCCGCCGGCG
SpeI (107) 1 site
ACTAGTTGATCA
SalI (119) 1 site
GTCGACCAGCTG
SgrDI (119) 1 site
CGTCGACGGCAGCTGC
AccI (120) 1 site
GTMKACCAKMTG
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.Sticky ends from different AccI sites may not be compatible.
EcoRV (127) 1 site
GATATCCTATAG
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
TspMI (130) 1 site
CCCGGGGGGCCC
XmaI (130) 1 site
CCCGGGGGGCCC
Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI (132) 1 site
CCCGGGGGGCCC
SmaI can be used at 37°C for brief incubations.
EcoRI (135) 1 site
GAATTCCTTAAG
RsrII (142) 1 site
CGGWCCGGCCWGGC
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.For full activity, add fresh DTT.
AgeI (144) 1 site
ACCGGTTGGCCA
Acc65I (147) 1 site
GGTACCCCATGG
KpnI (151) 1 site
GGTACCCCATGG
StuI (199) 1 site
AGGCCTTCCGGA
SacII (246) 1 site
CCGCGGGGCGCC
Efficient cleavage requires at least two copies of the SacII recognition sequence.
AvrII (249) 1 site
CCTAGGGGATCC
BbsI (278) 1 site
GAAGACNNCTTCTGNN(N)4
Sticky ends from different BbsI sites may not be compatible.BbsI gradually loses activity when stored at -20°C.
BsmBI (323) 1 site
CGTCTCNGCAGAGN(N)4
Sticky ends from different BsmBI sites may not be compatible.
NcoI (552) 1 site
CCATGGGGTACC
SnaBI (576) 1 site
TACGTAATGCAT
NsiI (863) 1 site
ATGCATTACGTA
BstAPI (1079) 1 site
GCANNNNNTGCCGTNNNNNACG
Sticky ends from different BstAPI sites may not be compatible.
BspQI (1139) 1 site
GCTCTTCNCGAGAAGNNNN
Sticky ends from different BspQI sites may not be compatible.
SapI (1139) 1 site
GCTCTTCNCGAGAAGNNNN
Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PciI (1255) 1 site
ACATGTTGTACA
PciI is inhibited by nonionic detergents.
AlwNI (1671) 1 site
CAGNNNCTGGTCNNNGAC
Sticky ends from different AlwNI sites may not be compatible.
AhdI (2190) 1 site
GACNNNNNGTCCTGNNNNNCAG
The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible.
AmpR 2117 .. 2977 = 861 bp 286 amino acids = 31.6 kDa Segment 2: 2117 .. 2908 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 2117 .. 2977 = 861 bp 286 amino acids = 31.6 kDa Segment 1: signal sequence 2909 .. 2977 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 2117 .. 2977 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
ori 1316 .. 1904 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori 1316 .. 1904 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori 3265 .. 3720 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori 3265 .. 3720 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer 553 .. 856 = 304 bp human cytomegalovirus immediate early enhancer
CMV enhancer 553 .. 856 = 304 bp human cytomegalovirus immediate early enhancer
CMV promoter 349 .. 552 = 204 bp human cytomegalovirus (CMV) immediate early promoter
CMV promoter 349 .. 552 = 204 bp human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal 3846 .. 3980 = 135 bp SV40 polyadenylation signal
SV40 poly(A) signal 3846 .. 3980 = 135 bp SV40 polyadenylation signal
loxP 1982 .. 2015 = 34 bp Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP 1982 .. 2015 = 34 bp Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
attB2 36 .. 60 = 25 bp recombination site for the Gateway® BP reaction
attB2 36 .. 60 = 25 bp recombination site for the Gateway® BP reaction
attB1 153 .. 177 = 25 bp recombination site for the Gateway® BP reaction
attB1 153 .. 177 = 25 bp recombination site for the Gateway® BP reaction
T7 promoter 16 .. 34 = 19 bp promoter for bacteriophage T7 RNA polymerase
T7 promoter 16 .. 34 = 19 bp promoter for bacteriophage T7 RNA polymerase
SP6 promoter 178 .. 196 = 19 bp promoter for bacteriophage SP6 RNA polymerase
SP6 promoter 178 .. 196 = 19 bp promoter for bacteriophage SP6 RNA polymerase
M13 rev 207 .. 223 = 17 bp common sequencing primer, one of multiple similar variants
M13 rev 207 .. 223 = 17 bp common sequencing primer, one of multiple similar variants
M13 fwd 4385 .. 5 = 17 bp common sequencing primer, one of multiple similar variants
M13 fwd 4385 .. 5 = 17 bp common sequencing primer, one of multiple similar variants
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