Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV (3342) 1 site
GCCGGCCGGCCG
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
BtgZI (3242) 1 site
GCGATG(N)10CGCTAC(N)10(N)4
Sticky ends from different BtgZI sites may not be compatible.After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.BtgZI is typically used at 60°C, but is 75% active at 37°C.
PsiI (3113) 1 site
TTATAAAATATT
PfoI (2824) 1 site
TCCNGGAAGGNCCT
Sticky ends from different PfoI sites may not be compatible.
AatII (2713) 1 site
GACGTCCTGCAG
ZraI (2711) 1 site
GACGTCCTGCAG
ScaI (2271) 1 site
AGTACTTCATGA
NmeAIII (1939) 1 site
GCCGAG(N)18-19NNCGGCTC(N)18-19
Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM).
BpmI (1861) 1 site
CTGGAG(N)14NNGACCTC(N)14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.BpmI quickly loses activity at 37°C.
BsaI (1852) 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.
HindIII (183) 1 site
AAGCTTTTCGAA
PmlI (275) 1 site
CACGTGGTGCAC
PmlI gradually loses activity when stored at -20°C.
AarI (298) 1 site
CACCTGC(N)4GTGGACG(N)4(N)4
Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI (298) 1 site
ACCTGC(N)4TGGACG(N)4(N)4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI (298) 1 site
ACCTGC(N)4TGGACG(N)4(N)4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
Acc65I (400) 1 site
GGTACCCCATGG
KpnI (404) 1 site
GGTACCCCATGG
PflMI (412) 1 site
CCANNNNNTGGGGTNNNNNACC
Sticky ends from different PflMI sites may not be compatible.
BtgI (492) 1 site
CCRYGGGGYRCC
Sticky ends from different BtgI sites may not be compatible.
BmgBI (501) 1 site
CACGTCGTGCAG
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
MscI (566) 1 site
TGGCCAACCGGT
XcmI (574) 1 site
CCANNNNNNNNNTGGGGTNNNNNNNNNACC
The 1-base overhangs produced by XcmI may be hard to ligate.Sticky ends from different XcmI sites may not be compatible.
NdeI (576) 1 site
CATATGGTATAC
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BstXI (577) 1 site
CCANNNNNNTGGGGTNNNNNNACC
Sticky ends from different BstXI sites may not be compatible.
BamHI (581) 1 site
GGATCCCCTAGG
After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRI (587) 1 site
GAATTCCTTAAG
EcoRV (595) 1 site
GATATCCTATAG
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
PacI (603) 1 site
TTAATTAAAATTAATT
PstI (612) 1 site
CTGCAGGACGTC
Eco53kI (616) 1 site
GAGCTCCTCGAG
SacI (618) 1 site
GAGCTCCTCGAG
SalI (620) 1 site
GTCGACCAGCTG
SgrDI (620) 1 site
CGTCGACGGCAGCTGC
AccI (621) 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.
HincII (622) 1 site
GTYRACCARYTG
EagI (627) 1 site
CGGCCGGCCGGC
NotI (627) 1 site
GCGGCCGCCGCCGGCG
AvaI (635) 1 site
CYCGRGGRGCYC
Sticky ends from different AvaI sites may not be compatible.
BsoBI (635) 1 site
CYCGRGGRGCYC
Sticky ends from different BsoBI sites may not be compatible.BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I (635) 1 site
CTCGAGGAGCTC
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI (635) 1 site
VCTCGAGBBGAGCTCV
XhoI (635) 1 site
CTCGAGGAGCTC
BmeT110I (636) 1 site
CYCGRGGRGCYC
AleI (637) 1 site
CACNNNNGTGGTGNNNNCAC
BglII (643) 1 site
AGATCTTCTAGA
PmeI (687) 1 site
GTTTAAACCAAATTTG
SpeI (692) 1 site
ACTAGTTGATCA
BlpI (701) 1 site
GCTNAGCCGANTCG
Sticky ends from different BlpI sites may not be compatible.
BseYI (1202) 1 site
CCCAGCGGGTCG
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI (1206) 1 site
CCCAGCGGGTCG
AlwNI (1314) 1 site
CAGNNNCTGGTCNNNGAC
Sticky ends from different AlwNI sites may not be compatible.
AhdI (1791) 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 1718 .. 2578 = 861 bp 286 amino acids = 31.6 kDa Segment 2: 1718 .. 2509 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 1718 .. 2578 = 861 bp 286 amino acids = 31.6 kDa Segment 1: signal sequence 2510 .. 2578 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 1718 .. 2578 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
ori 959 .. 1547 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori 959 .. 1547 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES 70 .. 531 = 462 bp internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES 70 .. 531 = 462 bp internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
f1 ori 3017 .. 3472 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori 3017 .. 3472 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter 2579 .. 2683 = 105 bp
AmpR promoter 2579 .. 2683 = 105 bp
MCS 564 .. 642 = 79 bp multiple cloning site
MCS 564 .. 642 = 79 bp multiple cloning site
T7 terminator 712 .. 759 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator 712 .. 759 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase
poly(A) 654 .. 683 = 30 bp
poly(A) 654 .. 683 = 30 bp
FLAG 539 .. 562 = 24 bp 8 amino acids = 1.0 kDa Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG 539 .. 562 = 24 bp 8 amino acids = 1.0 kDa Product: FLAG® epitope tag, followed by an enterokinase cleavage site
T7 promoter 1 .. 19 = 19 bp promoter for bacteriophage T7 RNA polymerase
T7 promoter 1 .. 19 = 19 bp promoter for bacteriophage T7 RNA polymerase
ATG 533 .. 535 = 3 bp 1 amino acid = 149.2 Da Product: start codon
ATG 533 .. 535 = 3 bp 1 amino acid = 149.2 Da Product: start codon
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