pRS424
Yeast episomal vector with a TRP1 marker and an MCS derived from pBLUESCRIPT II.
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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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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. |
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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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Sticky ends from different BbsI sites may not be compatible.BbsI gradually loses activity when stored at -20°C. |
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Efficient cleavage requires at least two copies of the NmeAIII recognition sequence. Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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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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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 BstAPI sites may not be compatible. |
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Sticky ends from different Bsu36I sites may not be compatible. |
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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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Sticky ends from different BtgZI sites may not be compatible.After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.BtgZI is typically used at 60°C, but is 75% active at 37°C. |
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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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ApaI can be used between 25°C and 37°C. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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SmaI can be used at 37°C for brief incubations. |
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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 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 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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Sticky ends from different AflIII sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
| 2μ ori 4206 .. 5548 = 1343 bp yeast 2μ plasmid origin of replication |
| 2μ ori 4206 .. 5548 = 1343 bp yeast 2μ plasmid origin of replication |
| AmpR 3214 .. 4074 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 3214 .. 4005 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3214 .. 4074 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 4006 .. 4074 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3214 .. 4074 = 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 |
| ori 2455 .. 3043 = 589 bp high-copy-number colE1/pMB1/pBR322/pUC origin of replication |
| ori 2455 .. 3043 = 589 bp high-copy-number colE1/pMB1/pBR322/pUC origin of replication |
| lacZα 1479 .. 2057 = 579 bp 192 amino acids = 20.6 kDa Product: LacZα fragment of β-galactosidase |
| lacZα 1479 .. 2057 = 579 bp 192 amino acids = 20.6 kDa Product: LacZα fragment of β-galactosidase |
| TRP1 promoter 187 .. 467 = 281 bp |
| TRP1 promoter 187 .. 467 = 281 bp |
| AmpR promoter 4075 .. 4179 = 105 bp |
| AmpR promoter 4075 .. 4179 = 105 bp |
| lac promoter 2101 .. 2131 = 31 bp 3 segments Segment 3: -10 2101 .. 2107 = 7 bp promoter for the E. coli lac operon |
| lac promoter 2101 .. 2131 = 31 bp 3 segments Segment 2: 2108 .. 2125 = 18 bp promoter for the E. coli lac operon |
| lac promoter 2101 .. 2131 = 31 bp 3 segments Segment 1: -35 2126 .. 2131 = 6 bp promoter for the E. coli lac operon |
| lac promoter 2101 .. 2131 = 31 bp 3 segments promoter for the E. coli lac operon |
| lac operator 2077 .. 2093 = 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 2077 .. 2093 = 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). |
| 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 |
| MCS 1894 .. 2001 = 108 bp pBluescript multiple cloning site |
| MCS 1894 .. 2001 = 108 bp pBluescript multiple cloning site |
| T7 promoter 1867 .. 1885 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 1867 .. 1885 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T3 promoter 2014 .. 2032 = 19 bp promoter for bacteriophage T3 RNA polymerase |
| T3 promoter 2014 .. 2032 = 19 bp promoter for bacteriophage T3 RNA polymerase |
| M13 fwd 1841 .. 1857 = 17 bp common sequencing primer, one of multiple similar variants |
| M13 fwd 1841 .. 1857 = 17 bp common sequencing primer, one of multiple similar variants |
| M13 rev 2053 .. 2069 = 17 bp common sequencing primer, one of multiple similar variants |
| M13 rev 2053 .. 2069 = 17 bp common sequencing primer, one of multiple similar variants |
| ORF: 718 .. 999 = 282 bp ORF: 93 amino acids = 10.8 kDa |
| ORF: 3344 .. 3610 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 4241 .. 4531 = 291 bp ORF: 96 amino acids = 11.5 kDa |
| ORF: 468 .. 1142 = 675 bp ORF: 224 amino acids = 24.1 kDa |
| ORF: 3214 .. 4074 = 861 bp ORF: 286 amino acids = 31.6 kDa |
| ORF: 1479 .. 2057 = 579 bp ORF: 192 amino acids = 20.6 kDa |
| ORF: 5007 .. 5327 = 321 bp ORF: 106 amino acids = 12.9 kDa |
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