Plasmid Files

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PciI  (4181) 1 site

PciI is inhibited by nonionic detergents.

ApaLI  (3867) 1 site

BsaI  (3252) 1 site

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

PfoI  (3038) 1 site

Sticky ends from different PfoI sites may not be compatible.

BstBI  (2945) 1 site

RsrII  (2779) 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.

BsrDI  (2496) 1 site

Sticky ends from different BsrDI sites may not be compatible.

PflFI  (2381) 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  (2381) 1 site

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

FspI  (2365) 1 site

MscI  (2345) 1 site

PluTI  (2266) 1 site

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

SfoI  (2264) 1 site

NarI  (2263) 1 site

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

KasI  (2262) 1 site

BsaBI  (2122) 1 site

* Blocked by Dam methylation.

NdeI  (240) 1 site

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

SnaBI  (346) 1 site

NheI  (597) 1 site

BmtI  (601) 1 site

Eco53kI  (653) 1 site

SacI  (655) 1 site

AleI  (661) 1 site

SacII  (662) 1 site

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

BstXI  (663) 1 site

Sticky ends from different BstXI sites may not be compatible.

NotI  (668) 1 site

SrfI  (771) 1 site

BamHI  (777) 1 site

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

PstI  (793) 1 site

EcoRI  (795) 1 site

EcoRV  (803) 1 site

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

HindIII  (807) 1 site

SalI  (822) 1 site

AccI  (823) 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  (824) 1 site

AbsI  (828) 1 site

PaeR7I  (828) 1 site

PaeR7I does not recognize the sequence CTCTCGAG.

PspXI  (828) 1 site

XhoI  (828) 1 site

PspOMI  (837) 1 site

ApaI  (841) 1 site

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

PvuI  (919) 1 site

BclI  (928) 1 site

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

BbsI  (1130) 1 site

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

MluI  (1149) 1 site

PsiI  (1251) 1 site

DraIII  (1379) 1 site

Sticky ends from different DraIII sites may not be compatible.

SfiI  (2038) 1 site

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

BseRI  (2081) 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.

StuI  (2084) 1 site

NeoR/KanR 2135 .. 2929  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin)

NeoR/KanR 2135 .. 2929  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin)

ori 3537 .. 4125  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 3537 .. 4125  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

f1 ori 1155 .. 1610  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

f1 ori 1155 .. 1610  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

SV40 promoter 1743 .. 2100  =  358 bp SV40 enhancer and early promoter

SV40 promoter 1743 .. 2100  =  358 bp SV40 enhancer and early promoter

CMV enhancer 66 .. 370  =  305 bp human cytomegalovirus immediate early enhancer

CMV enhancer 66 .. 370  =  305 bp human cytomegalovirus immediate early enhancer

bGH poly(A) signal 941 .. 1148  =  208 bp bovine growth hormone polyadenylation signal

bGH poly(A) signal 941 .. 1148  =  208 bp bovine growth hormone polyadenylation signal

CMV promoter 371 .. 574  =  204 bp human cytomegalovirus (CMV) immediate early promoter

CMV promoter 371 .. 574  =  204 bp human cytomegalovirus (CMV) immediate early promoter

AmpR promoter 1637 .. 1739  =  103 bp

AmpR promoter 1637 .. 1739  =  103 bp

3xMyc 678 .. 767  =  90 bp 30 amino acids  =  3.6 kDa Product: 3 tandem Myc epitope tags

3xMyc 678 .. 767  =  90 bp 30 amino acids  =  3.6 kDa Product: 3 tandem Myc epitope tags

MCS 768 .. 842  =  75 bp multiple cloning site

MCS 768 .. 842  =  75 bp multiple cloning site

HSV TK poly(A) signal 3161 .. 3208  =  48 bp herpesvirus thymidine kinase polyadenylation signal

HSV TK poly(A) signal 3161 .. 3208  =  48 bp herpesvirus thymidine kinase polyadenylation signal

T3 promoter 620 .. 638  =  19 bp promoter for bacteriophage T3 RNA polymerase

T3 promoter 620 .. 638  =  19 bp promoter for bacteriophage T3 RNA polymerase

T7 promoter 888 .. 906  =  19 bp promoter for bacteriophage T7 RNA polymerase

T7 promoter 888 .. 906  =  19 bp promoter for bacteriophage T7 RNA polymerase

stop codons 850 .. 860  =  11 bp stop codons in all three reading frames

stop codons 850 .. 860  =  11 bp stop codons in all three reading frames

ATG 675 .. 677  =  3 bp 1 amino acid  =  149.2 Da Product: start codon

ATG 675 .. 677  =  3 bp 1 amino acid  =  149.2 Da Product: start codon

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