pCDFDuet-1
Bacterial vector with a CloDF13 (CDF) 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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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 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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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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FseI gradually loses activity when stored at -20°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 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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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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Sticky ends from different BglI sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
| ACYCDuetUP1 Primer 19-mer / 63% GC 1 binding site 3722 .. 3740 = 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 2559 .. 3641 = 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 2559 .. 3641 = 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 680 .. 1471 = 792 bp 263 amino acids = 29.3 kDa Product: aminoglycoside adenylyltransferase confers resistance to spectinomycin and streptomycin |
| SmR 680 .. 1471 = 792 bp 263 amino acids = 29.3 kDa Product: aminoglycoside adenylyltransferase confers resistance to spectinomycin and streptomycin |
| CloDF13 ori 1611 .. 2349 = 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 1611 .. 2349 = 739 bp Plasmids containing the CloDF13 (CDF) 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 1472 .. 1563 = 92 bp |
| AmpR promoter 1472 .. 1563 = 92 bp |
| lacI promoter 3642 .. 3719 = 78 bp |
| lacI promoter 3642 .. 3719 = 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 3765 .. 2 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 3765 .. 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 |
| ATG 300 .. 302 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 300 .. 302 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| S-Tag 366 .. 410 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A |
| S-Tag 366 .. 410 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A |
| ATG 71 .. 73 = 3 bp 1 amino acid = 149.2 Da |
| ATG 71 .. 73 = 3 bp 1 amino acid = 149.2 Da |
| 6xHis 83 .. 100 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 83 .. 100 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| ORF: 1049 .. 1432 = 384 bp ORF: 127 amino acids = 14.6 kDa |
| ORF: 2525 .. 2776 = 252 bp ORF: 83 amino acids = 9.1 kDa |
| ORF: 894 .. 1172 = 279 bp ORF: 92 amino acids = 9.4 kDa |
| ORF: 1182 .. 1439 = 258 bp ORF: 85 amino acids = 9.1 kDa |
| ORF: 680 .. 1471 = 792 bp ORF: 263 amino acids = 29.3 kDa |
| ORF: 2539 .. 2802 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 3391 .. 158 = 549 bp ORF: 182 amino acids = 19.6 kDa |
| ORF: 2559 .. 3518 = 960 bp ORF: 319 amino acids = 34.1 kDa |
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