pMCSG30
Bacterial expression vector with a 6xHis-TEV-MBP leader and a SacB negative selection marker, for high-throughput purification of recombinant proteins.
Sequence Author: Midwest Center for Structural Genomics
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Sticky ends from different AvaI sites may not be compatible. |
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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. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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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 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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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 PpuMI sites may not be compatible. |
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PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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ApaI can be used between 25°C and 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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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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Efficient cleavage requires at least two copies of the SfiI recognition sequence. Sticky ends from different SfiI 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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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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| * Blocked by Dcm methylation. |
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Efficient cleavage requires at least two copies of the SfiI recognition sequence. Sticky ends from different SfiI sites may not be compatible. |
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Sticky ends from different BsmI 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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BsiWI is typically used at 55°C, but is 50% active at 37°C. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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The 1-base overhangs produced by EcoNI may be hard to ligate.Sticky ends from different EcoNI sites may not be compatible. |
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Sticky ends from different PflMI sites may not be compatible. |
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| SacB 255 .. 1676 = 1422 bp 473 amino acids = 53.0 kDa 2 segments Segment 2: 255 .. 1589 = 1335 bp 444 amino acids = 50.0 kDa Product: secreted levansucrase that renders bacterial growth sensitive to sucrose negative selection marker |
| SacB 255 .. 1676 = 1422 bp 473 amino acids = 53.0 kDa 2 segments Segment 1: signal peptide 1590 .. 1676 = 87 bp 29 amino acids = 3.0 kDa Product: secreted levansucrase that renders bacterial growth sensitive to sucrose negative selection marker |
| SacB 255 .. 1676 = 1422 bp 473 amino acids = 53.0 kDa 2 segments Product: secreted levansucrase that renders bacterial growth sensitive to sucrose negative selection marker |
| MBP 2171 .. 3244 = 1074 bp 358 amino acids = 39.4 kDa Product: maltose binding protein from E. coli This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol. |
| MBP 2171 .. 3244 = 1074 bp 358 amino acids = 39.4 kDa Product: maltose binding protein from E. coli This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol. |
| TEV site 3251 .. 3271 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 3251 .. 3271 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| 6xHis 3296 .. 3313 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 3296 .. 3313 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| ATG 3314 .. 3316 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 3314 .. 3316 = 3 bp 1 amino acid = 149.2 Da |