pMCSG24 (linearized)
Linearized bacterial coexpression vector for ligation-independent cloning (LIC), with a 6xHis-GST-TEV leader and a CloDF13 origin.
Sequence Author: Midwest Center for Structural Genomics
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Sticky ends from different PfoI sites may not be compatible. |
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Sticky ends from different BstAPI 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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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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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 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. |
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| lacI 2262 .. 3344 = 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 2262 .. 3344 = 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 383 .. 1174 = 792 bp 263 amino acids = 29.3 kDa Product: aminoglycoside adenylyltransferase (Murphy, 1985) confers resistance to spectinomycin and streptomycin |
| SmR 383 .. 1174 = 792 bp 263 amino acids = 29.3 kDa Product: aminoglycoside adenylyltransferase (Murphy, 1985) confers resistance to spectinomycin and streptomycin |
| ATG 3784 .. 3786 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 3784 .. 3786 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| 6xHis 3787 .. 3804 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 3787 .. 3804 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| GST 3823 .. 4476 = 654 bp 218 amino acids = 25.5 kDa Product: glutathione S-transferase from Schistosoma japonicum |
| GST 3823 .. 4476 = 654 bp 218 amino acids = 25.5 kDa Product: glutathione S-transferase from Schistosoma japonicum |
| TEV site 4504 .. 4524 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 4504 .. 4524 = 21 bp 7 amino acids = 900.0 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| CloDF13 ori 1314 .. 2052 = 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 1314 .. 2052 = 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 1175 .. 1266 = 92 bp |
| AmpR promoter 1175 .. 1266 = 92 bp |
| lacI promoter 3345 .. 3422 = 78 bp |
| lacI promoter 3345 .. 3422 = 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 3555 .. 3557 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 3555 .. 3557 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| 6xHis 3567 .. 3584 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 3567 .. 3584 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| lac operator 3717 .. 3741 = 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 3717 .. 3741 = 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 3487 .. 3511 = 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 3487 .. 3511 = 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 3698 .. 3716 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3698 .. 3716 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3468 .. 3486 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3468 .. 3486 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| RBS 3542 .. 3547 = 6 bp ribosome binding site |
| RBS 3542 .. 3547 = 6 bp ribosome binding site |
| RBS 3770 .. 3775 = 6 bp ribosome binding site |
| RBS 3770 .. 3775 = 6 bp ribosome binding site |
| ORF: 3784 .. 4527 = 744 bp ORF: 248 amino acids = 29.0 kDa |
| ORF: 752 .. 1135 = 384 bp ORF: 127 amino acids = 14.6 kDa |
| ORF: 2228 .. 2479 = 252 bp ORF: 83 amino acids = 9.1 kDa |
| ORF: 597 .. 875 = 279 bp ORF: 92 amino acids = 9.4 kDa |
| ORF: 885 .. 1142 = 258 bp ORF: 85 amino acids = 9.1 kDa |
| ORF: 2242 .. 2505 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 3094 .. 3642 = 549 bp ORF: 182 amino acids = 19.6 kDa |
| ORF: 2262 .. 3221 = 960 bp ORF: 319 amino acids = 34.1 kDa |
| ORF: 383 .. 1174 = 792 bp ORF: 263 amino acids = 29.3 kDa |
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