Plasmid Files

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SgrDI  (7709) 1 site

PvuI  (7158) 1 site

PciI  (5895) 1 site

PciI is inhibited by nonionic detergents.

PfoI  (5319) 1 site

Sticky ends from different PfoI sites may not be compatible.

RsrII  (5058) 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.

PflFI  (4660) 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  (4660) 1 site

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

PluTI  (4545) 1 site

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

SfoI  (4543) 1 site

NarI  (4542) 1 site

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

KasI  (4541) 1 site

EagI  (4448) 1 site

BclI  (4383) 1 site

* Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 37°C.

AvrII  (4332) 1 site

StuI  (4331) 1 site

SexAI  (4099) 1 site

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

MfeI  (161) 1 site

SpeI  (249) 1 site

NdeI  (484) 1 site

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

SnaBI  (590) 1 site

Eco53kI  (816) 1 site

SacI  (818) 1 site

BstXI  (1572) 1 site

Sticky ends from different BstXI sites may not be compatible.

BmgBI  (1651) 1 site

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.

SrfI  (1685) 1 site

BbvCI  (1828) 1 site

BamHI  (2099) 1 site

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

EcoRI  (2552) 1 site

BspEI  (2556) 1 site

XbaI  (3179) 1 site

* Blocked by Dam methylation.

PspOMI  (3185) 1 site

ApaI  (3189) 1 site

ApaI can be used between 25°C and 37°C.

SacII  (3192) 1 site

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

BstBI  (3196) 1 site

AgeI  (3246) 1 site

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

PmeI  (3264) 1 site

DraIII  (3809) 1 site

Sticky ends from different DraIII sites may not be compatible.

AmpR 6715 .. 7575  =  861 bp 286 amino acids  =  31.6 kDa    Segment 2:      6715 .. 7506  =  792 bp    263 amino acids  =  28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 6715 .. 7575  =  861 bp 286 amino acids  =  31.6 kDa    Segment 1:  signal sequence      7507 .. 7575  =  69 bp    23 amino acids  =  2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 6715 .. 7575  =  861 bp 286 amino acids  =  31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

NeoR/KanR 4414 .. 5208  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®)

NeoR/KanR 4414 .. 5208  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®)

CmR 2111 .. 2770  =  660 bp 219 amino acids  =  25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol

CmR 2111 .. 2770  =  660 bp 219 amino acids  =  25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol

ori 5956 .. 6544  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 5956 .. 6544  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

f1 ori 3576 .. 4004  =  429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

f1 ori 3576 .. 4004  =  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 4018 .. 4347  =  330 bp SV40 enhancer and early promoter

SV40 promoter 4018 .. 4347  =  330 bp SV40 enhancer and early promoter

ccdB 1464 .. 1769  =  306 bp 101 amino acids  =  11.7 kDa Product: CcdB, a bacterial toxin that poisons DNA gyrase Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.

ccdB 1464 .. 1769  =  306 bp 101 amino acids  =  11.7 kDa Product: CcdB, a bacterial toxin that poisons DNA gyrase Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.

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

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

attR1 911 .. 1035  =  125 bp recombination site for the Gateway® LR reaction

attR1 911 .. 1035  =  125 bp recombination site for the Gateway® LR reaction

attR2 3051 .. 3175  =  125 bp recombination site for the Gateway® LR reaction

attR2 3051 .. 3175  =  125 bp recombination site for the Gateway® LR reaction

SV40 poly(A) signal 5384 .. 5505  =  122 bp SV40 polyadenylation signal

SV40 poly(A) signal 5384 .. 5505  =  122 bp SV40 polyadenylation signal

AmpR promoter 7576 .. 7680  =  105 bp

AmpR promoter 7576 .. 7680  =  105 bp

cat promoter 2771 .. 2873  =  103 bp promoter of the E. coli cat gene

cat promoter 2771 .. 2873  =  103 bp promoter of the E. coli cat gene

HSV TK poly(A) signal 3326 .. 3374  =  49 bp herpesvirus thymidine kinase polyadenylation signal

HSV TK poly(A) signal 3326 .. 3374  =  49 bp herpesvirus thymidine kinase polyadenylation signal

V5 tag 3201 .. 3242  =  42 bp 14 amino acids  =  1.4 kDa Product: epitope tag from simian virus 5

V5 tag 3201 .. 3242  =  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

SV40 ori 4198 .. 4333  =  136 bp SV40 origin of replication

SV40 ori 4198 .. 4333  =  136 bp SV40 origin of replication

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