pMAL-p5X
Bacterial vector for inducible periplasmic expression of maltose-binding protein (MBP) fusions with a Factor Xa cleavage site.
Sequence Author: New England Biolabs
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| * Blocked by Dcm methylation. |
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Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present. This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.Sticky ends from different Bpu10I 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 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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PciI is inhibited by nonionic detergents. |
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Sticky ends from different Bsu36I sites may not be compatible. |
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Sticky ends from different BglI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the RsrII recognition sequence. Sticky ends from different RsrII sites may not be compatible.For full activity, add fresh DTT. |
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Sticky ends from different PflMI sites may not be compatible. |
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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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| * 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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BsiWI is typically used at 55°C, but is 50% active at 37°C. |
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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 BsmI sites may not be compatible. |
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Sticky ends from different BlpI sites may not be compatible. |
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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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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Sticky ends from different StyI 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 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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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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| MBP 1528 .. 2703 = 1176 bp 392 amino acids = 42.9 kDa 2 segments Segment 1: signal sequence 1528 .. 1605 = 78 bp 26 amino acids = 2.7 kDa Product: maltose binding protein from E. coli This gene encodes a signal sequence that can direct MBP to the bacterial periplasm. |
| MBP 1528 .. 2703 = 1176 bp 392 amino acids = 42.9 kDa 2 segments Segment 2: 1606 .. 2703 = 1098 bp 366 amino acids = 40.2 kDa Product: maltose binding protein from E. coli This gene encodes a signal sequence that can direct MBP to the bacterial periplasm. |
| MBP 1528 .. 2703 = 1176 bp 392 amino acids = 42.9 kDa 2 segments Product: maltose binding protein from E. coli This gene encodes a signal sequence that can direct MBP to the bacterial periplasm. |
| Factor Xa site 2752 .. 2763 = 12 bp 4 amino acids = 473.5 Da Product: Factor Xa recognition and cleavage site |
| Factor Xa site 2752 .. 2763 = 12 bp 4 amino acids = 473.5 Da Product: Factor Xa recognition and cleavage site |
| lacI 81 .. 1163 = 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 81 .. 1163 = 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 3162 .. 4022 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 3162 .. 3230 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3162 .. 4022 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 3231 .. 4022 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3162 .. 4022 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 4110 .. 4698 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 4110 .. 4698 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| rop 5068 .. 5259 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| rop 5068 .. 5259 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| AmpR promoter 3071 .. 3161 = 91 bp |
| AmpR promoter 3071 .. 3161 = 91 bp |
| rrnB T1 terminator 2838 .. 2924 = 87 bp transcription terminator T1 from the E. coli rrnB gene |
| rrnB T1 terminator 2838 .. 2924 = 87 bp transcription terminator T1 from the E. coli rrnB gene |
| lacIq promoter 3 .. 80 = 78 bp In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold. |
| lacIq promoter 3 .. 80 = 78 bp In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold. |
| tac promoter 1406 .. 1434 = 29 bp 3 segments Segment 1: -35 1406 .. 1411 = 6 bp strong E. coli promoter; hybrid between the trp and lac UV5 promoters |
| tac promoter 1406 .. 1434 = 29 bp 3 segments Segment 2: 1412 .. 1427 = 16 bp strong E. coli promoter; hybrid between the trp and lac UV5 promoters |
| tac promoter 1406 .. 1434 = 29 bp 3 segments Segment 3: -10 1428 .. 1434 = 7 bp strong E. coli promoter; hybrid between the trp and lac UV5 promoters |
| tac promoter 1406 .. 1434 = 29 bp 3 segments strong E. coli promoter; hybrid between the trp and lac UV5 promoters |
| rrnB T2 terminator 3016 .. 3043 = 28 bp transcription terminator T2 from the E. coli rrnB gene |
| rrnB T2 terminator 3016 .. 3043 = 28 bp transcription terminator T2 from the E. coli rrnB gene |
| lac operator 1442 .. 1458 = 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 1442 .. 1458 = 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). |
| MCS 2759 .. 2837 = 79 bp multiple cloning site |
| MCS 2759 .. 2837 = 79 bp multiple cloning site |
| ORF: 1528 .. 2829 = 1302 bp ORF: 433 amino acids = 47.2 kDa |