Duet Vectors
The Duet vectors are designed for the cloning and coexpression of two target ORFs. Every Duet vector contains two multiple cloning sites (MCS) each of which is preceded by a T7lac promoter and ribosome binding site (rbs). Multiple Duet vectors may be used together in compatible host strains to coexpress up to 8 target proteins (see detailed tables below).
- pACYCDuet-1 carries the P15A replicon, lacI gene and chloramphenicol resistance gene.
- pCDFDuet-1 carries the CloDF13 replicon, lacI gene and streptomycin/spectinomycin resistance gene (aadA).
- pCOLADuet-1 carries the ColA replicon, lacI gene and kanamycin resistance gene.
- pETDuet-1 carries the pBR322-derived ColE1 replicon, lacI gene and ampicillin resistance gene.
- pRSFDuet-1 carries the RSF1030 replicon, lacI gene and kanamycin resistance gene.
The vector sequences are provided in GenBank format, with significant features indicated. To convert the sequence into other formats, first select the sequence and copy (Command-C on Mac or Ctrl-C on Windows). Next, click here to open the NIH ReadSeq web page in a new window. Paste the sequence into the input field (Command-V on Mac or Ctrl-V on PC), choose your favorite format from the dropdown menu, and hit the "submit" button.
You can access sequences, maps, ordering information, newsletter articles and technical bulletins through the following links:
| Vector | Sequence | Map | Ordering Information | Newsletter Articles | Technical Bulletins |
|---|---|---|---|---|---|
| pACYCDuet-1 |  |
 |
 |
 |
 |
| pETDuet-1 | |
|
|
|
|
| pCDFDuet-1 | |
|
|
 |
|
| pCOLADuet-1 | |
|
|
 |
|
| pRSFDuet-1 | |
|
|
|
|
| Compatible Vector Combinations | Target Proteins (number) | Compatible Host Strain Group | |||
|---|---|---|---|---|---|
| Vector 1 | Vector 2 | Vector 3 | Vector 4 | ||
| pETDuet™-1 (AmpR) | pACYCDuet-1 (CamR) | pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
pCDFDuet-1 (SmR) | 8 | A |
| pETDuet-1 (AmpR) | pACYCDuet-1 (CamR) | pCDFDuet-1 (SmR) | none | 6 | B |
| pETDuet-1 (AmpR) | pACYCDuet-1 (CamR) | pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
none | 6 | A |
| pETDuet-1 (AmpR) | pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
pCDFDuet-1 (SmR) | none | 6 | C |
| pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
pACYCDuet-1 (CamR) | pCDFDuet-1 (SmR) | none | 6 | A |
| pETDuet-1 (AmpR) | pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
none | none | 4 | C |
| pACYCDuet-1 (CamR) | pETDuet-1 (AmpR) | none | none | 4 | B |
| pETDuet-1 (AmpR) | pCDFDuet-1 (SmR) | none | none | 4 | D |
| pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
pCDFDuet-1 (SmR) | none | none | 4 | C |
| pACYCDuet-1 (CamR) | pRSFDuet-1 (KanR) or pCOLADuet-1 (KanR) |
none | none | 4 | A |
| pACYCDuet-1 (CamR) | pCDFDuet-1 (SmR) | none | none | 4 | B |
| Resistance markers: AmpR: ampicillin/carbenicillin, KanR: kanamycin, CamR: chloramphenicol, SmR: streptomycin/spectinomycin | |||||
| Host Strain Groups | |||
|---|---|---|---|
| Group A B834(DE3) BL21(DE3) BLR(DE3) HMS174(DE3) Novablue(DE3) Origami 2(DE3)* Tuner™(DE3) |
Group C B834(DE3) B834(DE3)pLysS BL21(DE3) BL21(DE3)pLysS BLR(DE3) BLR(DE3)pLysS HMS174(DE3) HMS174(DE3)pLysS NovaBlue(DE3) Origami 2(DE3)* Origami 2(DE3)pLysS* Rosetta(DE3) Rosetta(DE3)pLysS Rosetta 2(DE3) Rosetta 2(DE3)pLysS RosettaBlue(DE3) RosettaBlue(DE3)pLysS Rosetta-gami 2(DE3)* Rosetta-gami 2(DE3)pLysS* Tuner(DE3) Tuner(DE3)pLysS |
Group D B834(DE3) B834(DE3)pLysS BL21(DE3) BL21(DE3)pLysS BLR(DE3) BLR(DE3)pLysS HMS174(DE3) HMS174(DE3)pLysS NovaBlue(DE3) Origami(DE3)* Origami(DE3)pLysS* Origami 2(DE3)* Origami 2(DE3)pLysS* Origami B(DE3) Origami B(DE3)pLysS Rosetta(DE3) Rosetta(DE3)pLysS Rosetta 2(DE3) Rosetta 2(DE3)pLysS RosettaBlue(DE3) RosettaBlue(DE3)pLysS Rosetta-gami(DE3)* Rosetta-gami(DE3)pLysS* Rosetta-gami 2(DE3)* Rosetta-gami 2(DE3)pLysS* Rosetta-gami B(DE3) Rosetta-gami B(DE3)pLysS Tuner(DE3) Tuner(DE3)pLysS |
|
| Group B B834(DE3) BL21(DE3) BLR(DE3) HMS174(DE3) NovaBlue(DE3) Origami(DE3)* Origami B(DE3) Origami 2(DE3)* Tuner(DE3) |
|||
| *These strains carry the rpsL mutation that confers resistance to streptomycin; therefore, spectinomycin must be used for selection of pCDFDuet recombinants. Resistance markers: AmpR, ampicillin/carbenicillin (50 µg/ml); KanR, kanamycin (30 µg/ml); CamR, chloramphenicol (34 µg/ml); SmR, streptomycin/spectinomycin (50 µg/ml). Note: the antibiotic concentrations used for four-way transformation plating and growth were reduced to 0.5 times the concentrations shown. |
|||
