pLATE11 (linearized)
Vector for ligation independent cloning (LIC) and tightly regulated bacterial expression of an untagged protein.
Sequence Author: Thermo Fisher
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Sticky ends from different BglI sites may not be compatible. |
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The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible. |
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Sticky ends from different AlwNI sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
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Sticky ends from different BspQI sites may not be compatible. |
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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. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.Sticky ends from different AccI sites may not be compatible. |
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The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible. |
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The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible. |
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Sticky ends from different PfoI sites may not be compatible. |
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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. |
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Efficient cleavage requires at least two copies of the BspMI recognition sequence. Sticky ends from different BspMI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the PaqCI recognition sequence. Sticky ends from different PaqCI sites may not be compatible.Cleavage can be improved with PaqCI Activator. |
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SwaI is typically used at 25°C, but is 50% active at 37°C. |
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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different StyI sites may not be compatible. |
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Sticky ends from different PflMI sites may not be compatible. |
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| * Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 37°C. |
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Sticky ends from different BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C. |
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ApaI can be used between 25°C and 37°C. |
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ApoI is typically used at 50°C, but is 50% active at 37°C. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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| * Blocked by Dcm methylation. Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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Efficient cleavage requires at least two copies of the PluTI recognition sequence. |
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| * Blocked by Dam methylation. |
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| * Blocked by Dam methylation. |
| LIC reverse sequencing primer 24-mer / 46% GC 3 binding sites 4297 .. 4312 = 16 annealed bases (38% GC) Tm = 48°C |
| LIC forward sequencing primer 20-mer / 40% GC 1 binding site 4360 .. 4379 = 20 annealed bases Tm = 49°C |
| LIC reverse sequencing primer 24-mer / 46% GC 3 binding sites 4387 .. 4401 = 15 annealed bases (40% GC) Tm = 47°C |
| LIC reverse sequencing primer 24-mer / 46% GC 3 binding sites 62 .. 85 = 24 annealed bases Tm = 58°C |
| lacI 296 .. 1378 = 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 296 .. 1378 = 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). |
| AmpR 3009 .. 3869 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 3009 .. 3800 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3009 .. 3869 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 3801 .. 3869 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3009 .. 3869 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 2250 .. 2838 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 2250 .. 2838 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| rop 1631 .. 1822 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| rop 1631 .. 1822 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| AmpR promoter 3870 .. 3974 = 105 bp |
| AmpR promoter 3870 .. 3974 = 105 bp |
| rrnB T1 terminator 4075 .. 4156 = 82 bp transcription terminator T1 from the E. coli rrnB gene |
| rrnB T1 terminator 4075 .. 4156 = 82 bp transcription terminator T1 from the E. coli rrnB gene |
| lacI promoter 218 .. 295 = 78 bp |
| lacI promoter 218 .. 295 = 78 bp |
| T7 terminator 162 .. 209 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 162 .. 209 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| tet promoter 119 .. 147 = 29 bp 3 segments Segment 3: -10 119 .. 124 = 6 bp E. coli promoter for tetracycline efflux protein gene |
| tet promoter 119 .. 147 = 29 bp 3 segments Segment 2: 125 .. 141 = 17 bp E. coli promoter for tetracycline efflux protein gene |
| tet promoter 119 .. 147 = 29 bp 3 segments Segment 1: -35 142 .. 147 = 6 bp E. coli promoter for tetracycline efflux protein gene |
| tet promoter 119 .. 147 = 29 bp 3 segments E. coli promoter for tetracycline efflux protein gene |
| rrnB T2 terminator 4248 .. 4275 = 28 bp transcription terminator T2 from the E. coli rrnB gene |
| rrnB T2 terminator 4248 .. 4275 = 28 bp transcription terminator T2 from the E. coli rrnB gene |
| lac operator 4379 .. 4403 = 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 4379 .. 4403 = 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). |
| T7 promoter 4360 .. 4378 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 4360 .. 4378 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| lac operator 4293 .. 4309 = 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 4293 .. 4309 = 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). |
| RBS 4440 .. 4446 = 7 bp ribosome binding site |
| RBS 4440 .. 4446 = 7 bp ribosome binding site |
| ORF: 229 .. 546 = 318 bp ORF: 105 amino acids = 11.2 kDa |
| ORF: 1135 .. 1527 = 393 bp ORF: 130 amino acids = 14.0 kDa |
| ORF: |