Plasmid Files

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SgrDI  (5496) 1 site

SspI  (5379) 1 site

ScaI  (5055) 1 site

PvuI  (4945) 1 site

AhdI  (4575) 1 site

The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible.

PciI  (3682) 1 site

PciI is inhibited by nonionic detergents.

BstZ17I  (3303) 1 site

BsmI  (3251) 1 site

Sticky ends from different BsmI sites may not be compatible.

AanI  (3190) 2 sites

PfoI  (3106) 1 site

Sticky ends from different PfoI sites may not be compatible.

RsrII  (2845) 1 site

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.

BglII  (12) 1 site

MfeI  (161) 1 site

Bpu10I  (180) 1 site

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.

NdeI  (484) 1 site

Prolonged incubation with NdeI may lead to removal of additional nucleotides.

SnaBI  (590) 1 site

HindIII  (902) 1 site

Acc65I  (908) 1 site

KpnI  (912) 1 site

BamHI  (920) 1 site

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.

BstXI  (939) 2 sites

Sticky ends from different BstXI sites may not be compatible.

EcoRI  (943) 1 site

EcoRV  (955) 1 site

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.

BstXI  (965) 2 sites

Sticky ends from different BstXI sites may not be compatible.

NotI  (970) 1 site

PaeR7I  (976) 1 site

PaeR7I does not recognize the sequence CTCTCGAG.

PspXI  (976) 1 site

XhoI  (976) 1 site

BstEII  (981) 1 site

Sticky ends from different BstEII sites may not be compatible. BstEII is typically used at 60°C, but is 50% active at 37°C.

BstBI  (991) 1 site

AgeI  (1041) 1 site

AgeI quickly loses activity at 37°C, but can be used at 25°C for long incubations.

PmeI  (1071) 1 site

BbsI  (1282) 1 site

Sticky ends from different BbsI sites may not be compatible. BbsI gradually loses activity when stored at -20°C.

DraIII  (1596) 1 site

Sticky ends from different DraIII sites may not be compatible.

AanI  (1721) 2 sites

SexAI  (1886) 1 site

* Blocked by Dcm methylation. Sticky ends from different SexAI sites may not be compatible.

StuI  (2118) 1 site

AvrII  (2119) 1 site

TspMI  (2140) 1 site

XmaI  (2140) 1 site

Efficient cleavage requires at least two copies of the XmaI recognition sequence. Full cleavage with XmaI may require a long incubation.

SmaI  (2142) 1 site

SmaI can be used at 37°C for brief incubations.

BsaBI  (2188) 1 site

* Blocked by Dam methylation.

KasI  (2328) 1 site

NarI  (2329) 1 site

Efficient cleavage requires at least two copies of the NarI recognition sequence.

SfoI  (2330) 1 site

BbeI  (2332) 1 site

Efficient cleavage requires at least two copies of the BbeI recognition sequence.

MscI  (2411) 1 site

PflFI  (2447) 1 site

The 1-base overhangs produced by PflFI may be hard to ligate. Sticky ends from different PflFI sites may not be compatible.

Tth111I  (2447) 1 site

The 1-base overhangs produced by Tth111I may be hard to ligate. Sticky ends from different Tth111I sites may not be compatible.

BssHII  (2726) 1 site

BssHII is typically used at 50°C, but is 75% active at 37°C.

AmpR 4502 .. 5362  =  861 bp 286 amino acids  =  31.6 kDa    Segment 2:      4502 .. 5293  =  792 bp    263 amino acids  =  28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 4502 .. 5362  =  861 bp 286 amino acids  =  31.6 kDa    Segment 1:  signal sequence      5294 .. 5362  =  69 bp    23 amino acids  =  2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 4502 .. 5362  =  861 bp 286 amino acids  =  31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

NeoR/KanR 2201 .. 2995  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin)

NeoR/KanR 2201 .. 2995  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin)

ori 3743 .. 4331  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 3743 .. 4331  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

f1 ori 1363 .. 1791  =  429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

f1 ori 1363 .. 1791  =  429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

CMV enhancer 235 .. 614  =  380 bp human cytomegalovirus immediate early enhancer

CMV enhancer 235 .. 614  =  380 bp human cytomegalovirus immediate early enhancer

SV40 promoter 1805 .. 2134  =  330 bp SV40 enhancer and early promoter

SV40 promoter 1805 .. 2134  =  330 bp SV40 enhancer and early promoter

bGH poly(A) signal 1093 .. 1317  =  225 bp bovine growth hormone polyadenylation signal

bGH poly(A) signal 1093 .. 1317  =  225 bp bovine growth hormone polyadenylation signal

CMV promoter 615 .. 818  =  204 bp human cytomegalovirus (CMV) immediate early promoter

CMV promoter 615 .. 818  =  204 bp human cytomegalovirus (CMV) immediate early promoter

SV40 poly(A) signal 3171 .. 3292  =  122 bp SV40 polyadenylation signal

SV40 poly(A) signal 3171 .. 3292  =  122 bp SV40 polyadenylation signal

AmpR promoter 5363 .. 5467  =  105 bp

AmpR promoter 5363 .. 5467  =  105 bp

MCS 902 .. 995  =  94 bp multiple cloning site

MCS 902 .. 995  =  94 bp multiple cloning site

V5 tag 996 .. 1037  =  42 bp 14 amino acids  =  1.4 kDa Product: epitope tag from simian virus 5

V5 tag 996 .. 1037  =  42 bp 14 amino acids  =  1.4 kDa Product: epitope tag from simian virus 5

T7 promoter 863 .. 881  =  19 bp promoter for bacteriophage T7 RNA polymerase

T7 promoter 863 .. 881  =  19 bp promoter for bacteriophage T7 RNA polymerase

6xHis 1047 .. 1064  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

6xHis 1047 .. 1064  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

SV40 ori 1985 .. 2120  =  136 bp SV40 origin of replication

SV40 ori 1985 .. 2120  =  136 bp SV40 origin of replication

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