The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible.
BpmI (3284) 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.
BglI (3235) 1 site
GCCNNNNNGGCCGGNNNNNCCG
Sticky ends from different BglI sites may not be compatible.
ScaI (2872) 1 site
AGTACTTCATGA
XmnI (2753) 1 site
GAANNNNTTCCTTNNNNAAG
EarI (2554) 1 site
CTCTTCNGAGAAGNNNN
Efficient cleavage requires at least two copies of the EarI recognition sequence.
Sticky ends from different EarI sites may not be compatible.
BglII (2486) 1 site
AGATCTTCTAGA
SmaI (2482) 1 site
CCCGGGGGGCCC
SmaI can be used at 37°C for brief incubations.
TspMI (2480) 1 site
CCCGGGGGGCCC
XmaI (2480) 1 site
CCCGGGGGGCCC
Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
BmgBI (2477) 1 site
CACGTCGTGCAG
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
DraIII (2453) 1 site
CACNNNGTGGTGNNNCAC
Sticky ends from different DraIII sites may not be compatible.
BsaBI (2346) 1 site
GATNNNNATCCTANNNNTAG
* Blocked by Dam methylation.
NruI (2279) 1 site
TCGCGAAGCGCT
* Blocked by Dam methylation.
PvuII (2255) 1 site
CAGCTGGTCGAC
PpuMI (2207) 1 site
RGGWCCYYCCWGGR
Sticky ends from different PpuMI sites may not be compatible.
BbsI (2133) 1 site
GAAGACNNCTTCTGNN(N)4
Sticky ends from different BbsI sites may not be compatible.BbsI gradually loses activity when stored at -20°C.
MscI (2051) 1 site
TGGCCAACCGGT
NcoI (2046) 1 site
CCATGGGGTACC
StyI (2046) 1 site
CCWWGGGGWWCC
Sticky ends from different StyI sites may not be compatible.
HincII (1984) 1 site
GTYRACCARYTG
PmlI (1979) 1 site
CACGTGGTGCAC
PmlI gradually loses activity when stored at -20°C.
BspHI (0) 1 site
TCATGAAGTACT
BstZ17I (216) 1 site
GTATACCATATG
BsiWI (317) 1 site
CGTACGGCATGC
BsiWI is typically used at 55°C, but is 50% active at 37°C.
RsrII (335) 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.
KasI (474) 1 site
GGCGCCCCGCGG
NarI (475) 1 site
GGCGCCCCGCGG
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI (476) 1 site
GGCGCCCCGCGG
PluTI (478) 1 site
GGCGCCCCGCGG
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
HindIII (557) 1 site
AAGCTTTTCGAA
Acc65I (563) 1 site
GGTACCCCATGG
KpnI (567) 1 site
GGTACCCCATGG
Eco53kI (571) 1 site
GAGCTCCTCGAG
SacI (573) 1 site
GAGCTCCTCGAG
BamHI (575) 1 site
GGATCCCCTAGG
After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
SpeI (581) 1 site
ACTAGTTGATCA
EcoRI (598) 1 site
GAATTCCTTAAG
EcoRV (610) 1 site
GATATCCTATAG
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
NotI (625) 1 site
GCGGCCGCCGCCGGCG
PaeR7I (631) 1 site
CTCGAGGAGCTC
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI (631) 1 site
VCTCGAGBBGAGCTCV
XhoI (631) 1 site
CTCGAGGAGCTC
XbaI (637) 1 site
TCTAGAAGATCT
SacII (650) 1 site
CCGCGGGGCGCC
Efficient cleavage requires at least two copies of the SacII recognition sequence.
AgeI (704) 1 site
ACCGGTTGGCCA
PsiI (811) 1 site
TTATAAAATATT
BspDI (878) 1 site
ATCGATTAGCTA
* Blocked by Dam methylation.
ClaI (878) 1 site
ATCGATTAGCTA
* Blocked by Dam methylation.
PciI (941) 1 site
ACATGTTGTACA
PciI is inhibited by nonionic detergents.
BciVI (1144) 1 site
GTATCC(N)5NCATAGG(N)5
The 1-base overhangs produced by BciVI may be hard to ligate.Sticky ends from different BciVI sites may not be compatible.
AmpR 2566 .. 3426 = 861 bp 286 amino acids = 31.6 kDa Segment 1: signal sequence 2566 .. 2634 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 2566 .. 3426 = 861 bp 286 amino acids = 31.6 kDa Segment 2: 2635 .. 3426 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR 2566 .. 3426 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics
ori 1002 .. 1590 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori 1002 .. 1590 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
OpIE-2 promoter 1 .. 548 = 548 bp strong constitutive baculovirus promoter for insect cell expression
OpIE-2 promoter 1 .. 548 = 548 bp strong constitutive baculovirus promoter for insect cell expression
BSD 2048 .. 2446 = 399 bp 132 amino acids = 13.7 kDa Product: blasticidin S deaminase confers resistance to blasticidin
BSD 2048 .. 2446 = 399 bp 132 amino acids = 13.7 kDa Product: blasticidin S deaminase confers resistance to blasticidin
OpIE-1 promoter 1665 .. 1956 = 292 bp moderate constitutive baculovirus promoter for insect cell expression
OpIE-1 promoter 1665 .. 1956 = 292 bp moderate constitutive baculovirus promoter for insect cell expression
OpIE-2 poly(A) signal 745 .. 874 = 130 bp baculovirus polyadenylation signal
OpIE-2 poly(A) signal 745 .. 874 = 130 bp baculovirus polyadenylation signal
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