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 Product: start codon |
lacI 3795 .. 4877 = 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 3795 .. 4877 = 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 7066 .. 7926 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 7066 .. 7857 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 7066 .. 7926 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 7858 .. 7926 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 7066 .. 7926 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
ori 6307 .. 6895 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 6307 .. 6895 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
sacB promoter 1677 .. 2122 = 446 bp sacB promoter and control region |
sacB promoter 1677 .. 2122 = 446 bp sacB promoter and control region |
rop 5686 .. 5877 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
rop 5686 .. 5877 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
AmpR promoter 7927 .. 8030 = 104 bp |
AmpR promoter 7927 .. 8030 = 104 bp |
lacI promoter 3717 .. 3794 = 78 bp |
lacI promoter 3717 .. 3794 = 78 bp |
T7 terminator 26 .. 73 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
T7 terminator 26 .. 73 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
lac operator 3361 .. 3385 = 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 3361 .. 3385 = 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 3386 .. 3404 = 19 bp promoter for bacteriophage T7 RNA polymerase |
T7 promoter 3386 .. 3404 = 19 bp promoter for bacteriophage T7 RNA polymerase |
6xHis 140 .. 157 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
6xHis 140 .. 157 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
RBS 3325 .. 3330 = 6 bp ribosome binding site |
RBS 3325 .. 3330 = 6 bp ribosome binding site |
ORF: 4933 .. 5289 = 357 bp ORF: 118 amino acids = 13.0 kDa |
ORF: 5653 .. 5877 = 225 bp ORF: 74 amino acids = 8.5 kDa |
ORF: 3128 .. 4045 = 918 bp ORF: 305 amino acids = 32.8 kDa |
ORF: 4634 .. 4897 = 264 bp ORF: 87 amino acids = 8.9 kDa |
ORF: 7196 .. 7462 = 267 bp ORF: 88 amino acids = 9.2 kDa |
ORF: 3918 .. 4877 = 960 bp ORF: 319 amino acids = 34.1 kDa |
ORF: 2123 .. 3316 = 1194 bp ORF: 397 amino acids = 43.4 kDa |
ORF: 3368 .. 3619 = 252 bp ORF: 83 amino acids = 8.6 kDa |
ORF: 3493 .. 3756 = 264 bp ORF: 87 amino acids = 9.5 kDa |
ORF: 4660 .. 4911 = 252 bp ORF: 83 amino acids = 9.1 kDa |
ORF: 7066 .. 7926 = 861 bp ORF: 286 amino acids = 31.5 kDa |
ORF: 255 .. 1676 = 1422 bp ORF: 473 amino acids = 53.0 kDa |
ORF: 5286 .. 5654 = 369 bp ORF: 122 amino acids = 14.2 kDa |
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