Plasmid Files

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PluTI  (3832) 1 site

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

SfoI  (3830) 1 site

NarI  (3829) 1 site

* Blocked by Dcm methylation. Efficient cleavage requires at least two copies of the NarI recognition sequence.

KasI  (3828) 1 site

EcoRV  (3639) 1 site

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

ApaI  (3400) 1 site

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

BanII  (3400) 1 site

Sticky ends from different BanII sites may not be compatible.

PspOMI  (3396) 1 site

BstEII  (3370) 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.

BclI  (3203) 1 site

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

MluI  (3189) 1 site

PflMI  (2771) 1 site

Sticky ends from different PflMI sites may not be compatible.

NsiI  (2752) 1 site

SphI  (2750) 1 site

MauBI  (2728) 1 site

PfoI  (2660) 1 site

Sticky ends from different PfoI sites may not be compatible.

PflFI  (2561) 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  (2561) 1 site

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

BstZ17I  (2536) 1 site

NdeI  (2484) 1 site

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

BspQI  (2422) 1 site

Sticky ends from different BspQI sites may not be compatible.

SapI  (2422) 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.

PciI  (2305) 1 site

PciI is inhibited by nonionic detergents.

AlwNI  (1896) 1 site

Sticky ends from different AlwNI sites may not be compatible.

BfuAI  (4032) 1 site

Efficient cleavage requires at least two copies of the BfuAI recognition sequence. Sticky ends from different BfuAI sites may not be compatible. BfuAI is typically used at 50°C, but is 50% active at 37°C.

BspMI  (4032) 1 site

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

BtgI  (4371) 1 site

Sticky ends from different BtgI sites may not be compatible.

NcoI  (4371) 1 site

StyI  (4371) 1 site

Sticky ends from different StyI sites may not be compatible.

End  (4381) 0 sites

Start  (1) 0 sites

EcoRI  (7) 1 site

BstBI  (11) 1 site

HindIII  (14) 1 site

SnaBI  (21) 1 site

SalI  (64) 1 site

PmeI  (94) 1 site

ScaI  (936) 1 site

TsoI  (1019) 1 site

Sticky ends from different TsoI sites may not be compatible. After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility. For full activity, add fresh S-adenosylmethionine (SAM).

PvuI  (1048) 1 site

FspI  (1194) 1 site

BglI  (1299) 1 site

Sticky ends from different BglI sites may not be compatible.

AhdI  (1417) 1 site

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

lacI 2839 .. 3921  =  1083 bp 360 amino acids  =  38.5 kDa Product: lac repressor 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).

lacI 2839 .. 3921  =  1083 bp 360 amino acids  =  38.5 kDa Product: lac repressor 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).

AmpR 630 .. 1490  =  861 bp 286 amino acids  =  31.5 kDa    Segment 1:  signal sequence      630 .. 698  =  69 bp    23 amino acids  =  2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 630 .. 1490  =  861 bp 286 amino acids  =  31.5 kDa    Segment 2:      699 .. 1490  =  792 bp    263 amino acids  =  28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 630 .. 1490  =  861 bp 286 amino acids  =  31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

ori 1661 .. 2249  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 1661 .. 2249  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

AmpR promoter 538 .. 629  =  92 bp

AmpR promoter 538 .. 629  =  92 bp

rrnB T1 terminator 313 .. 399  =  87 bp transcription terminator T1 from the E. coli rrnB gene

rrnB T1 terminator 313 .. 399  =  87 bp transcription terminator T1 from the E. coli rrnB gene

lacIq promoter 2761 .. 2838  =  78 bp In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.

lacIq promoter 2761 .. 2838  =  78 bp In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.

rrnG antiterminator 4223 .. 4292  =  70 bp antiterminator from the E. coli rrnG leader region (Berg et al., 1989)

rrnG antiterminator 4223 .. 4292  =  70 bp antiterminator from the E. coli rrnG leader region (Berg et al., 1989)

Myc 25 .. 54  =  30 bp 10 amino acids  =  1.2 kDa Product: Myc (human c-Myc oncogene) epitope tag

Myc 25 .. 54  =  30 bp 10 amino acids  =  1.2 kDa Product: Myc (human c-Myc oncogene) epitope tag

trc promoter 4153 .. 4182  =  30 bp    Segment 1:  -35      4153 .. 4158  =  6 bp strong E. coli promoter; hybrid between the trp and lac UV5 promoters

trc promoter 4153 .. 4182  =  30 bp    Segment 2:      4159 .. 4175  =  17 bp strong E. coli promoter; hybrid between the trp and lac UV5 promoters

trc promoter 4153 .. 4182  =  30 bp    Segment 3:  -10      4176 .. 4182  =  7 bp strong E. coli promoter; hybrid between the trp and lac UV5 promoters

trc promoter 4153 .. 4182  =  30 bp 3 segments strong E. coli promoter; hybrid between the trp and lac UV5 promoters

rrnB T2 terminator 491 .. 518  =  28 bp transcription terminator T2 from the E. coli rrnB gene

rrnB T2 terminator 491 .. 518  =  28 bp transcription terminator T2 from the E. coli rrnB gene

minicistron 4343 .. 4369  =  27 bp 8 amino acids  =  1.1 kDa Product: synthetic cistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream cistron (Schoner, 1997) This first cistron should terminate 3 bp upstream of the ATG for the second cistron.

minicistron 4343 .. 4369  =  27 bp 8 amino acids  =  1.1 kDa Product: synthetic cistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream cistron (Schoner, 1997) This first cistron should terminate 3 bp upstream of the ATG for the second cistron.

6xHis 70 .. 87  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

6xHis 70 .. 87  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

lac operator 4190 .. 4206  =  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 4190 .. 4206  =  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).

gene 10 translational enhancer 4306 .. 4314  =  9 bp translational enhancer from gene 10 of bacteriophage T7 (Olins and Rangwala, 1989)

gene 10 translational enhancer 4306 .. 4314  =  9 bp translational enhancer from gene 10 of bacteriophage T7 (Olins and Rangwala, 1989)

RBS 4329 .. 4333  =  5 bp ribosome binding site

RBS 4329 .. 4333  =  5 bp ribosome binding site

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