pESC-TRP
Yeast episomal vector with a TRP1 marker, for galactose-regulated expression and tagging of up to two genes.
Sequence Author: Agilent Technologies
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The 1-base overhangs produced by XcmI may be hard to ligate.Sticky ends from different XcmI sites may not be compatible. |
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This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site. |
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Efficient cleavage requires at least two copies of the BpmI recognition sequence. Sticky ends from different BpmI sites may not be compatible.After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.BpmI quickly loses activity at 37°C. |
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Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°C. |
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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 PfoI sites may not be compatible. |
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| * Blocked by Dam methylation. |
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Sticky ends from different Bsu36I sites may not be compatible. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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Efficient cleavage requires at least two copies of the BsgI recognition sequence. Sticky ends from different BsgI sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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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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Sticky ends from different DraIII sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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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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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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ApaI can be used between 25°C and 37°C. |
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SmaI can be used at 37°C for brief incubations. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Sticky ends from different BtgI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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Sticky ends from different PpuMI sites may not be compatible. |
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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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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. |
| 2μ ori 5115 .. 6457 = 1343 bp yeast 2μ plasmid origin of replication |
| 2μ ori 5115 .. 6457 = 1343 bp yeast 2μ plasmid origin of replication |
| AmpR 4123 .. 4983 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 4123 .. 4914 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4123 .. 4983 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 4915 .. 4983 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4123 .. 4983 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| TRP1 468 .. 1142 = 675 bp 224 amino acids = 24.1 kDa Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis yeast auxotrophic marker |
| TRP1 468 .. 1142 = 675 bp 224 amino acids = 24.1 kDa Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis yeast auxotrophic marker |
| GAL1,10 promoter 2162 .. 2826 = 665 bp divergent inducible promoter, regulated by Gal4 |
| GAL1,10 promoter 2162 .. 2826 = 665 bp divergent inducible promoter, regulated by Gal4 |
| ori 3364 .. 3952 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 3364 .. 3952 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 1241 .. 1696 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 1241 .. 1696 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| TRP1 promoter 187 .. 467 = 281 bp |
| TRP1 promoter 187 .. 467 = 281 bp |
| CYC1 terminator 2932 .. 3121 = 190 bp transcription terminator for CYC1 |
| CYC1 terminator 2932 .. 3121 = 190 bp transcription terminator for CYC1 |
| ADH1 terminator 1779 .. 1944 = 166 bp transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene |
| ADH1 terminator 1779 .. 1944 = 166 bp transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene |
| AmpR promoter 4984 .. 5088 = 105 bp |
| AmpR promoter 4984 .. 5088 = 105 bp |
| MCS 2 2830 .. 2927 = 98 bp multiple cloning site 2 |
| MCS 2 2830 .. 2927 = 98 bp multiple cloning site 2 |
| MCS 1 2074 .. 2160 = 87 bp multiple cloning site 1 |
| MCS 1 2074 .. 2160 = 87 bp multiple cloning site 1 |
| UAS 2378 .. 2495 = 118 bp upstream activating sequence mediating Gal4-dependent induction |
| UAS 2378 .. 2495 = 118 bp upstream activating sequence mediating Gal4-dependent induction |
| Myc 2873 .. 2902 = 30 bp 10 amino acids = 1.2 kDa Product: Myc (human c-Myc oncogene) epitope tag |
| Myc 2873 .. 2902 = 30 bp 10 amino acids = 1.2 kDa Product: Myc (human c-Myc oncogene) epitope tag |
| FLAG 2092 .. 2115 = 24 bp 8 amino acids = 1.0 kDa Product: FLAG® epitope tag, followed by an enterokinase cleavage site |
| FLAG 2092 .. 2115 = 24 bp 8 amino acids = 1.0 kDa Product: FLAG® epitope tag, followed by an enterokinase cleavage site |
| ORF: 718 .. 999 = 282 bp ORF: 93 amino acids = 10.8 kDa |
| ORF: 4253 .. 4519 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 5150 .. 5440 = 291 bp ORF: 96 amino acids = 11.5 kDa |