pMCSG14 (linearized)
Linearized bacterial coexpression vector for ligation-independent cloning (LIC), with a 6xHis-GST-TEV leader and a p15A origin.
Sequence Author: Midwest Center for Structural Genomics
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Sticky ends from different BstAPI 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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The 1-base overhangs produced by BciVI may be hard to ligate.Sticky ends from different BciVI 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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Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present. Sticky ends from different AcuI sites may not be compatible.After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.For full activity, add fresh S-adenosylmethionine (SAM). |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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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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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 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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SwaI is typically used at 25°C, but is 50% active at 37°C. |
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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 BlpI 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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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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| lacI 2489 .. 3571 = 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 2489 .. 3571 = 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). |
| ATG 4011 .. 4013 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 4011 .. 4013 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| 6xHis 4014 .. 4031 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 4014 .. 4031 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| GST 4050 .. 4703 = 654 bp 218 amino acids = 25.5 kDa Product: glutathione S-transferase from Schistosoma japonicum |
| GST 4050 .. 4703 = 654 bp 218 amino acids = 25.5 kDa Product: glutathione S-transferase from Schistosoma japonicum |
| TEV site 4731 .. 4751 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 4731 .. 4751 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| CmR 432 .. 1091 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| CmR 432 .. 1091 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| p15A ori 1720 .. 2264 = 545 bp Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. |
| p15A ori 1720 .. 2264 = 545 bp Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. |
| cat promoter 1092 .. 1194 = 103 bp promoter of the E. coli cat gene |
| cat promoter 1092 .. 1194 = 103 bp promoter of the E. coli cat gene |
| lacI promoter 3572 .. 3649 = 78 bp |
| lacI promoter 3572 .. 3649 = 78 bp |
| T7 terminator 165 .. 212 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 165 .. 212 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| S-Tag 69 .. 113 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A |
| S-Tag 69 .. 113 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A |
| ATG 3782 .. 3784 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 3782 .. 3784 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| 6xHis 3794 .. 3811 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 3794 .. 3811 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| lac operator 3944 .. 3968 = 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 3944 .. 3968 = 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 3714 .. 3738 = 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 3714 .. 3738 = 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 3925 .. 3943 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3925 .. 3943 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3695 .. 3713 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3695 .. 3713 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| RBS 3769 .. 3774 = 6 bp ribosome binding site |
| RBS 3769 .. 3774 = 6 bp ribosome binding site |
| RBS 3997 .. 4002 = 6 bp ribosome binding site |
| RBS 3997 .. 4002 = 6 bp ribosome binding site |
| ORF: 2455 .. 2706 = 252 bp ORF: 83 amino acids = 9.1 kDa |
| ORF: 1635 .. 1913 = 279 bp ORF: 92 amino acids = 10.4 kDa |
| ORF: 4011 .. 4754 = 744 bp ORF: 248 amino acids = 29.0 kDa |
| ORF: 432 .. 1091 = 660 bp ORF: 219 amino acids = 25.7 kDa |
| ORF: 1602 .. 1874 = 273 bp ORF: 90 amino acids = 9.7 kDa |
| ORF: 2469 .. 2732 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 3321 .. 3869 = 549 bp ORF: 182 amino acids = 19.6 kDa |
| ORF: 2489 .. 3448 = 960 bp ORF: 319 amino acids = 34.1 kDa |
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