pMCSG62
Bacterial vector encoding tRNA genes for rare Arg and Ile codons, for expressing a protein with a 6xHis-AviTag™-TEV leader plus the BirA biotin ligase.
Sequence Author: Midwest Center for Structural Genomics
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Sticky ends from different BglI sites may not be compatible. |
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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 DrdI sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
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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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PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations. |
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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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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 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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PaeR7I does not recognize the sequence CTCTCGAG. |
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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 BsmI sites may not be compatible. |
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Sticky ends from different DraIII sites may not be compatible. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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| * Blocked by Dam methylation. |
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| * Blocked by Dam methylation. |
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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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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 PfoI sites may not be compatible. |
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Sticky ends from different PflMI sites may not be compatible. |
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Sticky ends from different BstAPI sites may not be compatible. |
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| * Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 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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| lacI 2351 .. 3433 = 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 2351 .. 3433 = 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). |
| BirA 241 .. 1206 = 966 bp 321 amino acids = 35.3 kDa Product: E. coli biotin ligase |
| BirA 241 .. 1206 = 966 bp 321 amino acids = 35.3 kDa Product: E. coli biotin ligase |
| AmpR 4621 .. 5481 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 4621 .. 5412 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4621 .. 5481 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 5413 .. 5481 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4621 .. 5481 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 3862 .. 4450 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 3862 .. 4450 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| AmpR promoter 5482 .. 5585 = 104 bp |
| AmpR promoter 5482 .. 5585 = 104 bp |
| TEV site 1251 .. 1271 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 1251 .. 1271 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| AviTag™ 1278 .. 1322 = 45 bp 15 amino acids = 1.8 kDa Product: peptide tag that allows for enzymatic biotinylation |
| AviTag™ 1278 .. 1322 = 45 bp 15 amino acids = 1.8 kDa Product: peptide tag that allows for enzymatic biotinylation |
| 6xHis 1329 .. 1346 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 1329 .. 1346 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| ATG 1347 .. 1349 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 1347 .. 1349 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| lacI promoter 2273 .. 2350 = 78 bp |
| lacI promoter 2273 .. 2350 = 78 bp |
| argU 1712 .. 1788 = 77 bp Product: E. coli tRNA-Arg recognizing AGA and AGG |
| argU 1712 .. 1788 = 77 bp Product: E. coli tRNA-Arg recognizing AGA and AGG |
| ileX 2058 .. 2133 = 76 bp Product: E. coli tRNA-Ile recognizing AUA |
| ileX 2058 .. 2133 = 76 bp Product: E. coli tRNA-Ile recognizing AUA |
| 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 1394 .. 1418 = 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 1394 .. 1418 = 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 1419 .. 1437 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 1419 .. 1437 = 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 1215 .. 1220 = 6 bp ribosome binding site |
| RBS 1215 .. 1220 = 6 bp ribosome binding site |
| RBS 1358 .. 1363 = 6 bp ribosome binding site |
| RBS 1358 .. 1363 = 6 bp ribosome binding site |
| ORF: 2134 .. 2601 = 468 bp ORF: 155 amino acids = 16.8 kDa |
| ORF: 3190 .. 3453 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 1847 .. 2101 = 255 bp ORF: 84 amino acids = 8.5 kDa |
| ORF: 2474 .. 3433 = 960 bp ORF: 319 amino acids = 34.1 kDa |
| ORF: 4751 .. 5017 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 1281 .. 1562 = 282 bp ORF: 93 amino acids = 10.5 kDa |
| ORF: 241 .. 1206 = 966 bp ORF: 321 amino acids = 35.3 kDa |
| ORF: 4621 .. 5481 = 861 bp ORF: 286 amino acids = 31.5 kDa |
| ORF: 3216 .. 3467 = 252 bp ORF: 83 amino acids = 9.1 kDa |
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