Sticky ends from different AflIII sites may not be compatible.
PciI (5385) 1 site
ACATGTTGTACA
PciI is inhibited by nonionic detergents.
ApaLI (5071) 1 site
GTGCACCACGTG
BsaI (4456) 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.
PfoI (4242) 1 site
TCCNGGAAGGNCCT
Sticky ends from different PfoI sites may not be compatible.
RsrII (3983) 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.
BsrDI (3700) 1 site
GCAATGNNCGTTAC
Sticky ends from different BsrDI sites may not be compatible.
PflFI (3585) 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 (3585) 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.
PluTI (3470) 1 site
GGCGCCCCGCGG
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI (3468) 1 site
GGCGCCCCGCGG
NarI (3467) 1 site
GGCGCCCCGCGG
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI (3466) 1 site
GGCGCCCCGCGG
BspDI (3307) 1 site
ATCGATTAGCTA
* Blocked by Dam methylation.
ClaI (3307) 1 site
ATCGATTAGCTA
* Blocked by Dam methylation.
SfiI (3242) 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.
AseI (7) 1 site
ATTAATTAATTA
NdeI (234) 1 site
CATATGGTATAC
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SnaBI (340) 1 site
TACGTAATGCAT
NheI (591) 1 site
GCTAGCCGATCG
BmtI (595) 1 site
GCTAGCCGATCG
BglII (609) 1 site
AGATCTTCTAGA
PaeR7I (613) 1 site
CTCGAGGAGCTC
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI (613) 1 site
CTCGAGGAGCTC
Eco53kI (618) 1 site
GAGCTCCTCGAG
SacI (620) 1 site
GAGCTCCTCGAG
HindIII (622) 1 site
AAGCTTTTCGAA
EcoRI (629) 1 site
GAATTCCTTAAG
SalI (639) 1 site
GTCGACCAGCTG
AccI (640) 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.
Acc65I (645) 1 site
GGTACCCCATGG
KpnI (649) 1 site
GGTACCCCATGG
SacII (652) 1 site
CCGCGGGGCGCC
Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspOMI (653) 1 site
GGGCCCCCCGGG
TspMI (656) 1 site
CCCGGGGGGCCC
XmaI (656) 1 site
CCCGGGGGGCCC
Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
ApaI (657) 1 site
GGGCCCCCCGGG
ApaI can be used between 25°C and 37°C.
SmaI (658) 1 site
CCCGGGGGGCCC
SmaI can be used at 37°C for brief incubations.
BamHI (660) 1 site
GGATCCCCTAGG
After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
AgeI (666) 1 site
ACCGGTTGGCCA
BsrGI (2099) 1 site
TGTACAACATGT
BsrGI is typically used at 37°C, but is even more active at 60°C.
NotI (2111) 1 site
GCGGCCGCCGCCGGCG
XbaI (2121) 1 site
TCTAGAAGATCT
* Blocked by Dam methylation.
MfeI (2217) 1 site
CAATTGGTTAAC
HpaI (2230) 1 site
GTTAACCAATTG
BtsαI (2306) 1 site
GCAGTGNNCGTCAC
Sticky ends from different BtsαI sites may not be compatible.
AflII (2349) 1 site
CTTAAGGAATTC
DraIII (2583) 1 site
CACNNNGTGGTGNNNCAC
Sticky ends from different DraIII sites may not be compatible.
tdTomato 679 .. 2109 = 1431 bp 476 amino acids = 54.2 kDa Product: tandem dimeric (pseudo-monomeric) derivative of DsRed mammalian codon-optimized
tdTomato 679 .. 2109 = 1431 bp 476 amino acids = 54.2 kDa Product: tandem dimeric (pseudo-monomeric) derivative of DsRed mammalian codon-optimized
NeoR/KanR 3339 .. 4133 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR 3339 .. 4133 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori 4741 .. 5329 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori 4741 .. 5329 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori 2359 .. 2814 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori 2359 .. 2814 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter 2947 .. 3304 = 358 bp SV40 enhancer and early promoter
SV40 promoter 2947 .. 3304 = 358 bp SV40 enhancer and early promoter
CMV enhancer 61 .. 364 = 304 bp human cytomegalovirus immediate early enhancer
CMV enhancer 61 .. 364 = 304 bp human cytomegalovirus immediate early enhancer
CMV promoter 365 .. 568 = 204 bp human cytomegalovirus (CMV) immediate early promoter
CMV promoter 365 .. 568 = 204 bp human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal 2231 .. 2352 = 122 bp SV40 polyadenylation signal
SV40 poly(A) signal 2231 .. 2352 = 122 bp SV40 polyadenylation signal
AmpR promoter 2841 .. 2945 = 105 bp
AmpR promoter 2841 .. 2945 = 105 bp
MCS 591 .. 671 = 81 bp multiple cloning site of fluorescent protein plasmids
MCS 591 .. 671 = 81 bp multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal 4365 .. 4412 = 48 bp herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal 4365 .. 4412 = 48 bp herpesvirus thymidine kinase polyadenylation signal
SV40 ori 3155 .. 3290 = 136 bp SV40 origin of replication
SV40 ori 3155 .. 3290 = 136 bp SV40 origin of replication