pMCSG75 (linearized)
Linearized bacterial vector for ligation-independent cloning (LIC), with a C-terminal TEV-AviTag™-6xHis cassette, the BirA biotin ligase, and tRNA genes for rare Arg and Ile codons.
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 BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 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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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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| * 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. |
| lacI 1002 .. 2084 = 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 1002 .. 2084 = 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 4761 .. 5726 = 966 bp 321 amino acids = 35.3 kDa Product: E. coli biotin ligase |
| BirA 4761 .. 5726 = 966 bp 321 amino acids = 35.3 kDa Product: E. coli biotin ligase |
| AmpR 3272 .. 4132 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 3272 .. 4063 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3272 .. 4132 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 4064 .. 4132 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3272 .. 4132 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 2513 .. 3101 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 2513 .. 3101 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| AmpR promoter 4133 .. 4236 = 104 bp |
| AmpR promoter 4133 .. 4236 = 104 bp |
| 6xHis 5778 .. 5795 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 5778 .. 5795 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| AviTag™ 5802 .. 5846 = 45 bp 15 amino acids = 1.8 kDa Product: peptide tag that allows for enzymatic biotinylation |
| AviTag™ 5802 .. 5846 = 45 bp 15 amino acids = 1.8 kDa Product: peptide tag that allows for enzymatic biotinylation |
| TEV site 5853 .. 5873 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 5853 .. 5873 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| lacI promoter 924 .. 1001 = 78 bp |
| lacI promoter 924 .. 1001 = 78 bp |
| argU 363 .. 439 = 77 bp Product: E. coli tRNA-Arg recognizing AGA and AGG |
| argU 363 .. 439 = 77 bp Product: E. coli tRNA-Arg recognizing AGA and AGG |
| ileX 709 .. 784 = 76 bp Product: E. coli tRNA-Ile recognizing AUA |
| ileX 709 .. 784 = 76 bp Product: E. coli tRNA-Ile recognizing AUA |
| T7 terminator 4546 .. 4593 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 4546 .. 4593 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| lac operator 45 .. 69 = 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 45 .. 69 = 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 70 .. 88 = 19 bp promoter for bacteriophage T7 RNA polymerase |