Plasmid Files

 
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PvuII  (15) 1 site

EcoO109I  (3997) 1 site

Sticky ends from different EcoO109I sites may not be compatible.

SspI  (3825) 1 site

ScaI  (3501) 1 site

PvuI  (3391) 1 site

FspI  (3243) 1 site

BpmI  (3091) 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.

BanI  (2969) 1 site

Sticky ends from different BanI sites may not be compatible.

AlwNI  (2544) 1 site

Sticky ends from different AlwNI sites may not be compatible.

AflIII  (2128) 1 site

Sticky ends from different AflIII sites may not be compatible.

PciI  (2128) 1 site

PciI is inhibited by nonionic detergents.

BsiWI  (25) 1 site

BsiWI is typically used at 55°C, but is 50% active at 37°C.

PstI  (35) 1 site

SalI  (37) 1 site

PsiI  (194) 1 site

AscI  (251) 1 site

BssHII  (251) 1 site

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

BbvCI  (257) 1 site

Bpu10I  (257) 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.

BamHI  (297) 1 site

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

PacI  (371) 1 site

BtgZI  (541) 1 site

Sticky ends from different BtgZI sites may not be compatible. After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility. BtgZI is typically used at 60°C, but is 75% active at 37°C.

AgeI  (767) 1 site

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

BseRI  (805) 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.

BglII  (930) 1 site

BsrGI  (935) 1 site

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

BspEI  (948) 1 site

* Blocked by Dam methylation.

PmlI  (1043) 1 site

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

XbaI  (1167) 1 site

MfeI  (1223) 1 site

Bsu36I  (1325) 1 site

Sticky ends from different Bsu36I sites may not be compatible.

BstXI  (1381) 1 site

Sticky ends from different BstXI sites may not be compatible.

BsgI  (1496) 1 site

Efficient cleavage requires at least two copies of the BsgI recognition sequence. Sticky ends from different BsgI sites may not be compatible. For full activity, add fresh S-adenosylmethionine (SAM).

AarI  (1516) 1 site

Efficient cleavage requires at least two copies of the AarI recognition sequence. Sticky ends from different AarI sites may not be compatible. After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.

PmeI  (1842) 1 site

Eco53kI  (1849) 1 site

SacI  (1851) 1 site

EcoRI  (1853) 1 site

BspDI  (1860) 1 site

ClaI  (1860) 1 site

SpeI  (1877) 1 site

SfiI  (1890) 1 site

Efficient cleavage requires at least two copies of the SfiI recognition sequence. Sticky ends from different SfiI sites may not be compatible.

BtgI  (1894) 1 site

Sticky ends from different BtgI sites may not be compatible.

SacII  (1897) 1 site

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

HpaI  (1949) 1 site

BspQI  (2012) 1 site

Sticky ends from different BspQI sites may not be compatible.

SapI  (2012) 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.

R3 20-mer  /  55% GC 1 binding site 261 .. 280  =  20 annealed bases Tm  =  58°C Reverse primer for promoter swapping and adding an N-terminal 3xHA tag. A gene-specific sequence should be added at the 5' end of the primer.

F4 20-mer  /  40% GC 1 binding site 1839 .. 1858  =  20 annealed bases Tm  =  52°C Forward primer for promoter swapping. This primer includes an EcoRI recognition sequence. A gene-specific sequence should be added at the 5' end of the primer.

AmpR 2948 .. 3808  =  861 bp 286 amino acids  =  31.6 kDa    Segment 2:      2948 .. 3739  =  792 bp    263 amino acids  =  28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 2948 .. 3808  =  861 bp 286 amino acids  =  31.6 kDa    Segment 1:  signal sequence      3740 .. 3808  =  69 bp    23 amino acids  =  2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 2948 .. 3808  =  861 bp 286 amino acids  =  31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

TRP1 1084 .. 1758  =  675 bp 224 amino acids  =  24.1 kDa Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis yeast auxotrophic marker

TRP1 1084 .. 1758  =  675 bp 224 amino acids  =  24.1 kDa Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis yeast auxotrophic marker

ori 2189 .. 2777  =  589 bp high-copy-number colE1/pMB1/pBR322/pUC origin of replication

ori 2189 .. 2777  =  589 bp high-copy-number colE1/pMB1/pBR322/pUC origin of replication

GAL1 promoter 400 .. 841  =  442 bp inducible promoter, regulated by Gal4

GAL1 promoter 400 .. 841  =  442 bp inducible promoter, regulated by Gal4

ADH1 terminator 50 .. 237  =  188 bp

ADH1 terminator 50 .. 237  =  188 bp

TRP1 promoter 936 .. 1083  =  148 bp

TRP1 promoter 936 .. 1083  =  148 bp

AmpR promoter 3809 .. 3913  =  105 bp

AmpR promoter 3809 .. 3913  =  105 bp

3xHA 270 .. 359  =  90 bp 30 amino acids  =  3.5 kDa Product: three tandem HA epitope tags

3xHA 270 .. 359  =  90 bp 30 amino acids  =  3.5 kDa Product: three tandem HA epitope tags

T7 promoter 1913 .. 1931  =  19 bp promoter for bacteriophage T7 RNA polymerase

T7 promoter 1913 .. 1931  =  19 bp promoter for bacteriophage T7 RNA polymerase

SP6 promoter 4259 .. 2  =  19 bp promoter for bacteriophage SP6 RNA polymerase

SP6 promoter 4259 .. 2  =  19 bp promoter for bacteriophage SP6 RNA polymerase

start codon 378 .. 380  =  3 bp 1 amino acid  =  149.2 Da

start codon 378 .. 380  =  3 bp 1 amino acid  =  149.2 Da

UAS 724 .. 841  =  118 bp upstream activating sequence mediating Gal4-dependent induction

UAS 724 .. 841  =  118 bp upstream activating sequence mediating Gal4-dependent induction

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