Plasmid Files

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BspQI  (2879) 1 site

Sticky ends from different BspQI sites may not be compatible.

SapI  (2879) 1 site

Sticky ends from different SapI sites may not be compatible. SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.

NspI  (2766) 1 site

PciI  (2762) 1 site

PciI is inhibited by nonionic detergents.

PspFI  (2462) 1 site

BseYI  (2458) 1 site

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.

AlwNI  (2353) 1 site

Sticky ends from different AlwNI sites may not be compatible.

AhdI  (1874) 1 site

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

BsaI  (1808) 1 site

Sticky ends from different BsaI sites may not be compatible. BsaI can be used between 37°C and 50°C.

BpmI  (1805) 1 site

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.

NmeAIII  (1727) 1 site

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).

MscI  (94) 1 site

BtgI  (98) 1 site

Sticky ends from different BtgI sites may not be compatible.

NcoI  (98) 1 site

BstBI  (134) 1 site

Acc65I  (157) 1 site

AgeI  (160) 1 site

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

KpnI  (161) 1 site

BseRI  (189) 1 site

Sticky ends from different BseRI sites may not be compatible. BseRI quickly loses activity at 37°C. Prolonged incubation with BseRI may lead to degradation of the DNA.

TspMI  (189) 1 site

XmaI  (189) 1 site

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

SmaI  (191) 1 site

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

SrfI  (191) 1 site

PstI  (212) 1 site

MluI  (214) 1 site

SnaBI  (222) 1 site

BamHI  (228) 1 site

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

EcoNI  (236) 1 site

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

EcoRI  (253) 1 site

EcoRV  (262) 1 site

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

SalI  (268) 1 site

AccI  (269) 1 site

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  (270) 1 site

HindIII  (274) 1 site

PaeR7I  (280) 1 site

PaeR7I does not recognize the sequence CTCTCGAG.

XhoI  (280) 1 site

AvrII  (286) 1 site

NheI  (291) 1 site

BmtI  (295) 1 site

XbaI  (297) 1 site

AleI  (308) 1 site

PmlI  (308) 1 site

PmlI gradually loses activity when stored at -20°C.

BstXI  (310) 1 site

Sticky ends from different BstXI sites may not be compatible.

EcoO109I  (315) 1 site

Sticky ends from different EcoO109I sites may not be compatible.

PspOMI  (315) 1 site

ApaI  (319) 1 site

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

Eco53kI  (323) 1 site

SacI  (325) 1 site

EagI  (328) 1 site

NotI  (328) 1 site

PsiI  (596) 1 site

DraIII  (724) 1 site

Sticky ends from different DraIII sites may not be compatible.

NgoMIV  (825) 1 site

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

NaeI  (827) 1 site

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

XmnI  (1274) 1 site

BsaHI  (1334) 1 site

BsaHI is typically used at 37°C, but is even more active at 60°C.

TatI  (1391) 1 site

ScaI  (1393) 1 site

AmpR 1087 .. 1947  =  861 bp 286 amino acids  =  31.6 kDa    Segment 1:  signal sequence      1087 .. 1155  =  69 bp    23 amino acids  =  2.6 kDa

AmpR 1087 .. 1947  =  861 bp 286 amino acids  =  31.6 kDa    Segment 2:      1156 .. 1947  =  792 bp    263 amino acids  =  28.9 kDa

AmpR 1087 .. 1947  =  861 bp 286 amino acids  =  31.6 kDa 2 segments

ori 2118 .. 2706  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 2118 .. 2706  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

LacZα 1 .. 549  =  549 bp 182 amino acids  =  20.4 kDa Product: LacZα fragment of β-galactosidase

LacZα 1 .. 549  =  549 bp 182 amino acids  =  20.4 kDa Product: LacZα fragment of β-galactosidase

AmpR promoter 982 .. 1086  =  105 bp

AmpR promoter 982 .. 1086  =  105 bp

lac promoter 3030 .. 3060  =  31 bp    Segment 1:  -35      3030 .. 3035  =  6 bp promoter for the E. coli lac operon

lac promoter 3030 .. 3060  =  31 bp    Segment 2:      3036 .. 3053  =  18 bp promoter for the E. coli lac operon

lac promoter 3030 .. 3060  =  31 bp    Segment 3:  -10      3054 .. 3060  =  7 bp promoter for the E. coli lac operon

lac promoter 3030 .. 3060  =  31 bp 3 segments promoter for the E. coli lac operon

lac operator 3068 .. 3084  =  17 bp The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).

lac operator 3068 .. 3084  =  17 bp The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).

f1 ori 500 .. 955  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

f1 ori 500 .. 955  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

MCS 92 .. 334  =  243 bp multiple cloning site

MCS 92 .. 334  =  243 bp multiple cloning site

T7 promoter 24 .. 42  =  19 bp promoter for bacteriophage T7 RNA polymerase

T7 promoter 24 .. 42  =  19 bp promoter for bacteriophage T7 RNA polymerase

M13 fwd 342 .. 358  =  17 bp common sequencing primer, one of multiple similar variants

M13 fwd 342 .. 358  =  17 bp common sequencing primer, one of multiple similar variants

M13 rev 3092 .. 5  =  17 bp common sequencing primer, one of multiple similar variants

M13 rev 3092 .. 5  =  17 bp common sequencing primer, one of multiple similar variants

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