Information
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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:

 

VectorSequenceMapOrdering
Information
Newsletter
Articles
Technical
Bulletins
pACYCDuet-1 Sequence Icon Map Icon Ordering Icon Newsletter 15 Icon Technical Bulletin Icon
pETDuet-1 Sequence Icon Map Icon Ordering Icon Newsletter 15 Icon Technical Bulletin Icon
pCDFDuet-1  Sequence Icon Map Icon Ordering Icon Newsletter 18 Icon Technical Bulletin Icon
pCOLADuet-1 Sequence Icon Map Icon Ordering Icon Newsletter 19 Icon Technical Bulletin Icon
pRSFDuet-1 Sequence Icon Map Icon Ordering Icon Newsletter 18 Icon Technical Bulletin Icon
Compatible Vector CombinationsTarget 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.

 

© Merck KGaA, Darmstadt, Germany, 2014


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