EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
ClaI (4965) 2 sites
ATCGATTAGCTA
BsaBI (4956) 1 site
GATNNNNATCCTANNNNTAG
* Blocked by Dam methylation.
PsiI (4835) 1 site
TTATAAAATATT
BssHII (4575) 1 site
GCGCGCCGCGCG
BssHII is typically used at 50°C, but is 75% active at 37°C.
EagI (4568) 1 site
CGGCCGGCCGGC
MscI (4495) 1 site
TGGCCAACCGGT
* Blocked by Dcm methylation.
BstEII (4254) 1 site
GGTNACCCCANTGG
Sticky ends from different BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII (4236) 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.
BsiWI (4176) 1 site
CGTACGGCATGC
BsiWI is typically used at 55°C, but is 50% active at 37°C.
SmaI (4171) 1 site
CCCGGGGGGCCC
SmaI can be used at 37°C for brief incubations.
TspMI (4169) 1 site
CCCGGGGGGCCC
XmaI (4169) 1 site
CCCGGGGGGCCC
Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
PflFI (4162) 1 site
GACNNNGTCCTGNNNCAG
The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible.
Tth111I (4162) 1 site
GACNNNGTCCTGNNNCAG
The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible.
PpuMI (4105) 1 site
RGGWCCYYCCWGGR
Sticky ends from different PpuMI sites may not be compatible.
AbsI (4043) 1 site
CCTCGAGGGGAGCTCC
PspXI (4043) 1 site
VCTCGAGBBGAGCTCV
AvrII (4021) 1 site
CCTAGGGGATCC
SfiI (3974) 1 site
GGCCNNNNNGGCCCCGGNNNNNCCGG
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
NcoI (3928) 1 site
CCATGGGGTACC
AlwNI (3283) 1 site
CAGNNNCTGGTCNNNGAC
Sticky ends from different AlwNI sites may not be compatible.
AhdI (2804) 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.
AseI (2629) 1 site
ATTAATTAATTA
EcoRI (1) 1 site
GAATTCCTTAAG
AgeI (89) 1 site
ACCGGTTGGCCA
BstXI (322) 1 site
CCANNNNNNTGGGGTNNNNNNACC
Sticky ends from different BstXI sites may not be compatible.
BsaAI (335) 1 site
YACGTRRTGCAY
AflII (394) 1 site
CTTAAGGAATTC
BglII (581) 1 site
AGATCTTCTAGA
PspOMI (628) 1 site
GGGCCCCCCGGG
ApaI (632) 1 site
GGGCCCCCCGGG
ApaI can be used between 25°C and 37°C.
NgoMIV (715) 1 site
GCCGGCCGGCCG
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI (717) 1 site
GCCGGCCGGCCG
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
FseI (719) 1 site
GGCCGGCCCCGGCCGG
FseI gradually loses activity when stored at -20°C.
Eco53kI (841) 1 site
GAGCTCCTCGAG
SacI (843) 1 site
GAGCTCCTCGAG
BbvCI (917) 1 site
CCTCAGCGGAGTCG
Bpu10I (917) 1 site
CCTNAGCGGANTCG
Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.Sticky ends from different Bpu10I sites may not be compatible.
BamHI (1195) 1 site
GGATCCCCTAGG
After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
XbaI (1201) 1 site
TCTAGAAGATCT
SbfI (1217) 1 site
CCTGCAGGGGACGTCC
HindIII (1225) 1 site
AAGCTTTTCGAA
EcoRV (1407) 2 sites
GATATCCTATAG
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
ClaI (1409) 2 sites
ATCGATTAGCTA
PvuII (1508) 1 site
CAGCTGGTCGAC
NdeI (1630) 1 site
CATATGGTATAC
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SspI (1999) 1 site
AATATTTTATAA
ScaI (2323) 1 site
AGTACTTCATGA
EF-1α promoter 12 .. 1193 = 1182 bp strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter 12 .. 1193 = 1182 bp strong constitutive promoter for human elongation factor EF-1α
AmpR 2017 .. 2877 = 861 bp 286 amino acids = 31.6 kDa Segment 1: signal sequence 2017 .. 2085 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 2017 .. 2877 = 861 bp 286 amino acids = 31.6 kDa Segment 2: 2086 .. 2877 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 2017 .. 2877 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR 4120 .. 4719 = 600 bp 199 amino acids = 21.5 kDa Product: puromycin N-acetyltransferase confers resistance to puromycin
PuroR 4120 .. 4719 = 600 bp 199 amino acids = 21.5 kDa Product: puromycin N-acetyltransferase confers resistance to puromycin
ori 3048 .. 3636 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori 3048 .. 3636 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter 3707 .. 4036 = 330 bp SV40 enhancer and early promoter
SV40 promoter 3707 .. 4036 = 330 bp SV40 enhancer and early promoter
SV40 poly(A) signal 4721 .. 4855 = 135 bp SV40 polyadenylation signal
SV40 poly(A) signal 4721 .. 4855 = 135 bp SV40 polyadenylation signal
AmpR promoter 1912 .. 2016 = 105 bp
AmpR promoter 1912 .. 2016 = 105 bp
HSV TK poly(A) signal 1288 .. 1335 = 48 bp herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal 1288 .. 1335 = 48 bp herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
M13 fwd 1422 .. 1438 = 17 bp common sequencing primer, one of multiple similar variants
M13 fwd 1422 .. 1438 = 17 bp common sequencing primer, one of multiple similar variants
EF-1α intron A 242 .. 1184 = 943 bp intron upstream of the start codon of human EF-1α
EF-1α intron A 242 .. 1184 = 943 bp intron upstream of the start codon of human EF-1α
SV40 ori 3887 .. 4022 = 136 bp SV40 origin of replication
SV40 ori 3887 .. 4022 = 136 bp SV40 origin of replication