Plasmid Files

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StyI  (57) 1 site

Sticky ends from different StyI sites may not be compatible.

ScaI  (5652) 1 site

BsaI  (5233) 1 site

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

AhdI  (5172) 1 site

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

PciI  (4279) 1 site

PciI is inhibited by nonionic detergents.

BspQI  (4163) 1 site

Sticky ends from different BspQI sites may not be compatible.

SapI  (4163) 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.

BstZ17I  (4050) 1 site

PflFI  (4024) 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  (4024) 1 site

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

PpuMI  (3285) 1 site

Sticky ends from different PpuMI sites may not be compatible.

FspAI  (3260) 1 site

BlpI  (80) 1 site

Sticky ends from different BlpI sites may not be compatible.

AvaI  (158) 1 site

Sticky ends from different AvaI sites may not be compatible.

BsoBI  (158) 1 site

Sticky ends from different BsoBI sites may not be compatible. BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.

PaeR7I  (158) 1 site

PaeR7I does not recognize the sequence CTCTCGAG.

PspXI  (158) 1 site

XhoI  (158) 1 site

BmeT110I  (159) 1 site

EagI  (166) 1 site

NotI  (166) 1 site

HindIII  (173) 1 site

SalI  (179) 1 site

Eco53kI  (188) 1 site

SacI  (190) 1 site

EcoRI  (192) 1 site

BamHI  (198) 1 site

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

BstBI  (446) 1 site

BsmI  (651) 1 site

Sticky ends from different BsmI sites may not be compatible.

DraIII  (1079) 1 site

Sticky ends from different DraIII sites may not be compatible.

SspI  (1247) 1 site

Acc65I  (1272) 1 site

KpnI  (1276) 1 site

NdeI  (1348) 1 site

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

XbaI  (1386) 1 site

BspDI  (1455) 1 site

* Blocked by Dam methylation.

ClaI  (1455) 1 site

* Blocked by Dam methylation.

SgrAI  (1497) 1 site

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

SphI  (1653) 1 site

EcoNI  (1713) 1 site

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

PflMI  (1760) 1 site

Sticky ends from different PflMI sites may not be compatible.

BstAPI  (1861) 1 site

Sticky ends from different BstAPI sites may not be compatible.

MluI  (2178) 1 site

BclI  (2192) 1 site

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

BstEII  (2359) 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.

PspOMI  (2385) 1 site

ApaI  (2389) 1 site

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

BssHII  (2589) 1 site

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

EcoRV  (2628) 1 site

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

HpaI  (2684) 1 site

PshAI  (3023) 1 site

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

lacI 1828 .. 2910  =  1083 bp 360 amino acids  =  38.6 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 1828 .. 2910  =  1083 bp 360 amino acids  =  38.6 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).

BirA 241 .. 1206  =  966 bp 321 amino acids  =  35.3 kDa Product: E. coli biotin ligase

BirA 241 .. 1206  =  966 bp 321 amino acids  =  35.3 kDa Product: E. coli biotin ligase

AmpR 5099 .. 5959  =  861 bp 286 amino acids  =  31.5 kDa    Segment 2:      5099 .. 5890  =  792 bp    263 amino acids  =  28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 5099 .. 5959  =  861 bp 286 amino acids  =  31.5 kDa    Segment 1:  signal sequence      5891 .. 5959  =  69 bp    23 amino acids  =  2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

AmpR 5099 .. 5959  =  861 bp 286 amino acids  =  31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics

ori 4340 .. 4928  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 4340 .. 4928  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

rop 3719 .. 3910  =  192 bp 63 amino acids  =  7.2 kDa Product: Rop protein, which maintains plasmids at low copy number

rop 3719 .. 3910  =  192 bp 63 amino acids  =  7.2 kDa Product: Rop protein, which maintains plasmids at low copy number

AmpR promoter 5960 .. 6063  =  104 bp

AmpR promoter 5960 .. 6063  =  104 bp

lacI promoter 1750 .. 1827  =  78 bp

lacI promoter 1750 .. 1827  =  78 bp

T7 terminator 26 .. 73  =  48 bp transcription terminator for bacteriophage T7 RNA polymerase

T7 terminator 26 .. 73  =  48 bp transcription terminator for bacteriophage T7 RNA polymerase

AviTag™ 1278 .. 1322  =  45 bp 15 amino acids  =  1.8 kDa Product: peptide tag that allows for enzymatic biotinylation

AviTag™ 1278 .. 1322  =  45 bp 15 amino acids  =  1.8 kDa Product: peptide tag that allows for enzymatic biotinylation

lac operator 1394 .. 1418  =  25 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 1394 .. 1418  =  25 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).

TEV site 1251 .. 1271  =  21 bp 7 amino acids  =  900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site

TEV site 1251 .. 1271  =  21 bp 7 amino acids  =  900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site

T7 promoter 1419 .. 1437  =  19 bp promoter for bacteriophage T7 RNA polymerase

T7 promoter 1419 .. 1437  =  19 bp promoter for bacteriophage T7 RNA polymerase

6xHis 140 .. 157  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

6xHis 140 .. 157  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

6xHis 1329 .. 1346  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

6xHis 1329 .. 1346  =  18 bp 6 amino acids  =  840.9 Da Product: 6xHis affinity tag

RBS 1215 .. 1220  =  6 bp ribosome binding site

RBS 1215 .. 1220  =  6 bp ribosome binding site

RBS 1358 .. 1363  =  6 bp ribosome binding site

RBS 1358 .. 1363  =  6 bp ribosome binding site

ATG 1347 .. 1349  =  3 bp 1 amino acid  =  149.2 Da Product: start codon

ATG 1347 .. 1349  =  3 bp 1 amino acid  =  149.2 Da Product: start codon

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