Plasmid Files

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AanI  (6559) 2 sites

ScaI  (5762) 1 site

AlwNI  (4805) 1 site

Sticky ends from different AlwNI sites may not be compatible.

PciI  (4389) 1 site

PciI is inhibited by nonionic detergents.

BstZ17I  (4010) 1 site

BsmI  (3958) 1 site

Sticky ends from different BsmI sites may not be compatible.

AanI  (3897) 2 sites

BstBI  (3720) 1 site

PflFI  (3156) 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  (3156) 1 site

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

NdeI  (592) 1 site

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

SnaBI  (698) 1 site

Eco53kI  (924) 1 site

SacI  (926) 1 site

EcoRI  (979) 1 site

BamHI  (992) 1 site

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

Acc65I  (998) 1 site

KpnI  (1002) 1 site

BmgBI  (1082) 1 site

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

BstEII  (1199) 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.

PasI  (1580) 1 site

Sticky ends from different PasI sites may not be compatible.

BspEI  (1734) 1 site

AsiSI  (1750) 1 site

SgfI  (1750) 1 site

MreI  (1752) 1 site

SgrAI  (1752) 1 site

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

AscI  (1756) 1 site

HindIII  (1770) 1 site

RsrII  (1787) 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.

MluI  (1793) 1 site

NotI  (1803) 1 site

PaeR7I  (1811) 1 site

PaeR7I does not recognize the sequence CTCTCGAG.

PspXI  (1811) 1 site

XhoI  (1811) 1 site

PmeI  (1820) 1 site

FseI  (1830) 1 site

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

BbsI  (2043) 1 site

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

EcoO109I  (2050) 1 site

Sticky ends from different EcoO109I sites may not be compatible.

BbvCI  (2146) 1 site

BlpI  (2190) 1 site

Sticky ends from different BlpI sites may not be compatible.

BsmBI  (2210) 1 site

Sticky ends from different BsmBI sites may not be compatible.

BstXI  (2297) 1 site

Sticky ends from different BstXI sites may not be compatible.

AgeI  (2408) 1 site

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

SexAI  (2595) 1 site

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

SfiI  (2781) 1 site

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

StuI  (2827) 1 site

AvrII  (2828) 1 site

BclI  (2879) 1 site

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

KasI  (3037) 1 site

NarI  (3038) 1 site

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

SfoI  (3039) 1 site

PluTI  (3041) 1 site

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

PstI  (3091) 1 site

VP1.5 (forward primer) 18-mer  /  44% GC 1 binding site 839 .. 856  =  18 annealed bases Tm  =  51°C

XL39 (reverse primer) 20-mer  /  55% GC 1 binding site 1926 .. 1945  =  20 annealed bases Tm  =  59°C

AmpR 5209 .. 6069  =  861 bp 286 amino acids  =  31.6 kDa    Segment 2:      5209 .. 6000  =  792 bp    263 amino acids  =  28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 5209 .. 6069  =  861 bp 286 amino acids  =  31.6 kDa    Segment 1:  signal sequence      6001 .. 6069  =  69 bp    23 amino acids  =  2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 5209 .. 6069  =  861 bp 286 amino acids  =  31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

NeoR/KanR 2910 .. 3704  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®)

NeoR/KanR 2910 .. 3704  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®)

TagCFP 1020 .. 1733  =  714 bp 238 amino acids  =  26.7 kDa Product: monomeric cyan fluorescent protein mammalian codon-optimized

TagCFP 1020 .. 1733  =  714 bp 238 amino acids  =  26.7 kDa Product: monomeric cyan fluorescent protein mammalian codon-optimized

hGH poly(A) signal 1863 .. 2485  =  623 bp human growth hormone polyadenylation signal

hGH poly(A) signal 1863 .. 2485  =  623 bp human growth hormone polyadenylation signal

ori 4450 .. 5038  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 4450 .. 5038  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

f1 ori 6201 .. 25  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

f1 ori 6201 .. 25  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

CMV enhancer 343 .. 722  =  380 bp human cytomegalovirus immediate early enhancer

CMV enhancer 343 .. 722  =  380 bp human cytomegalovirus immediate early enhancer

SV40 promoter 2514 .. 2843  =  330 bp SV40 enhancer and early promoter

SV40 promoter 2514 .. 2843  =  330 bp SV40 enhancer and early promoter

CMV promoter 723 .. 926  =  204 bp human cytomegalovirus (CMV) immediate early promoter

CMV promoter 723 .. 926  =  204 bp human cytomegalovirus (CMV) immediate early promoter

SV40 poly(A) signal 3878 .. 3999  =  122 bp SV40 polyadenylation signal

SV40 poly(A) signal 3878 .. 3999  =  122 bp SV40 polyadenylation signal

AmpR promoter 6070 .. 6174  =  105 bp

AmpR promoter 6070 .. 6174  =  105 bp

MCS 1734 .. 1832  =  99 bp multiple cloning site

MCS 1734 .. 1832  =  99 bp multiple cloning site

T7 promoter 952 .. 970  =  19 bp promoter for bacteriophage T7 RNA polymerase

T7 promoter 952 .. 970  =  19 bp promoter for bacteriophage T7 RNA polymerase

SV40 ori 2694 .. 2829  =  136 bp SV40 origin of replication

SV40 ori 2694 .. 2829  =  136 bp SV40 origin of replication

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