pRSFDuet-1
Bacterial vector with an RSF 1030 origin for the co-expression of two genes.
Sequence Author: MilliporeSigma (Novagen)
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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 BtgI 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 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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ApaI can be used between 25°C and 37°C. |
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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 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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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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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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FseI gradually loses activity when stored at -20°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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Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°C. |
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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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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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SmaI can be used at 37°C for brief incubations. |
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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. |
| ACYCDuetUP1 Primer 19-mer / 63% GC 1 binding site 3770 .. 3788 = 19 annealed bases Tm = 60°C |
| DuetUP2 Primer 20-mer / 50% GC 1 binding site 189 .. 208 = 20 annealed bases Tm = 57°C |
| DuetDOWN1 Primer 20-mer / 50% GC 1 binding site 189 .. 208 = 20 annealed bases Tm = 57°C |
| T7 Terminator Primer 19-mer / 53% GC 1 binding site 448 .. 466 = 19 annealed bases Tm = 57°C |
| lacI 2607 .. 3689 = 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 2607 .. 3689 = 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). |
| KanR 739 .. 1554 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin |
| KanR 739 .. 1554 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin |
| RSF ori 1662 .. 2411 = 750 bp Plasmids containing the RSF 1030 origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins. |
| RSF ori 1662 .. 2411 = 750 bp Plasmids containing the RSF 1030 origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins. |
| MCS-2 297 .. 438 = 142 bp multiple cloning site 2 |
| MCS-2 297 .. 438 = 142 bp multiple cloning site 2 |
| MCS-1 69 .. 168 = 100 bp multiple cloning site 1 |
| MCS-1 69 .. 168 = 100 bp multiple cloning site 1 |
| AmpR promoter 1555 .. 1646 = 92 bp |
| AmpR promoter 1555 .. 1646 = 92 bp |
| lacI promoter 3690 .. 3767 = 78 bp |
| lacI promoter 3690 .. 3767 = 78 bp |
| T7 terminator 462 .. 509 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 462 .. 509 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| 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 3813 .. 2 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3813 .. 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: 2573 .. 2824 = 252 bp ORF: 83 amino acids = 9.1 kDa |