pMCSG23
Bacterial coexpression vector with a 6xHis-MBP-TEV leader and a CloDF13 origin, for high-throughput purification of recombinant proteins.
Sequence Author: Midwest Center for Structural Genomics
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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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ApaI can be used between 25°C and 37°C. |
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Sticky ends from different BsmBI sites may not be compatible.BsmBI-v2 is an improved version of BsmBI. |
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Sticky ends from different Esp3I sites may not be compatible. |
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Efficient cleavage requires at least two copies of the PluTI recognition sequence. |
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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 SgrAI recognition sequence. |
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PciI is inhibited by nonionic detergents. |
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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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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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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 AlwNI 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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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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Sticky ends from different EcoO109I sites may not be compatible. |
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Sticky ends from different Bsu36I sites may not be compatible. |
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Sticky ends from different DrdI 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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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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Sticky ends from different BglI sites may not be compatible. |
| ATG 300 .. 302 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 300 .. 302 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| 6xHis 303 .. 320 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 303 .. 320 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| MBP 345 .. 1442 = 1098 bp 366 amino acids = 40.2 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 345 .. 1442 = 1098 bp 366 amino acids = 40.2 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 1464 .. 1484 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 1464 .. 1484 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| lacI 3749 .. 4831 = 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 3749 .. 4831 = 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). |
| SmR 1870 .. 2661 = 792 bp 263 amino acids = 29.3 kDa Product: aminoglycoside adenylyltransferase (Murphy, 1985) confers resistance to spectinomycin and streptomycin |
| SmR 1870 .. 2661 = 792 bp 263 amino acids = 29.3 kDa Product: aminoglycoside adenylyltransferase (Murphy, 1985) confers resistance to spectinomycin and streptomycin |
| CloDF13 ori 2801 .. 3539 = 739 bp Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins. |
| CloDF13 ori 2801 .. 3539 = 739 bp Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins. |
| AmpR promoter 2662 .. 2753 = 92 bp |
| AmpR promoter 2662 .. 2753 = 92 bp |
| lacI promoter 4832 .. 4909 = 78 bp |
| lacI promoter 4832 .. 4909 = 78 bp |
| T7 terminator 1652 .. 1699 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 1652 .. 1699 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| S-Tag 1556 .. 1600 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A |
| S-Tag 1556 .. 1600 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A |
| ATG 71 .. 73 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 71 .. 73 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| 6xHis 83 .. 100 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 83 .. 100 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| lac operator 233 .. 257 = 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 233 .. 257 = 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 3 .. 27 = 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 3 .. 27 = 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 4955 .. 2 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 4955 .. 2 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 214 .. 232 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 214 .. 232 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| RBS 58 .. 63 = 6 bp ribosome binding site |
| RBS 58 .. 63 = 6 bp ribosome binding site |
| RBS 286 .. 291 = 6 bp ribosome binding site |
| RBS 286 .. 291 = 6 bp ribosome binding site |
| ORF: 2239 .. 2622 = 384 bp ORF: 127 amino acids = 14.6 kDa |
| ORF: 3715 .. 3966 = 252 bp ORF: 83 amino acids = 9.1 kDa |
| ORF: 2084 .. 2362 = 279 bp ORF: 92 amino acids = 9.4 kDa |
| ORF: 2372 .. 2629 = 258 bp ORF: 85 amino acids = 9.1 kDa |
| ORF: 300 .. 1502 = 1203 bp ORF: 400 amino acids = 44.0 kDa |
| ORF: 1870 .. 2661 = 792 bp ORF: 263 amino acids = 29.3 kDa |
| ORF: 4581 .. 158 = 549 bp ORF: 182 amino acids = 19.6 kDa |
| ORF: 213 .. 461 = 249 bp ORF: 82 amino acids = 9.8 kDa |
| ORF: 3729 .. 3992 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 3749 .. 4708 = 960 bp ORF: 319 amino acids = 34.1 kDa |
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