Duet Expression Vectors

All vectors contain 2 MCS each preceded by a T7lac promoter and rbs. Vectors can be used in combination to express more than 2 proteins. See the Novagen web site for further details.

Vectors:

Vector MCS1 Tag MCS2 Tag Resistance* Replicon Sequence
pCDFDuet-1 NT His6 None Strep/Spect CloDF13 genbank
map
pCOLADuet-1 NT His6 None Kan ColA genbank
map
pETDuet-1 NT His6 None Amp ColE1 genbank
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pETDuetMOD1 NT His6, Tev cleavable None Amp ColE1 genbank
pETDuetMOD2 NT His6 NT His6, 3C cleavable Amp ColE1 genbank
pETDuetMOD3 NT His6, 3C cleavable None Amp ColE1 genbank
pRSFDuet-1 NT His6 None Kan RSF1030 genbank
map
pRSFDuetMOD1 NT His6, 3C cleavable None Kan RSF1030 genbank
*Amp=ampicillin; Kan=kanamycin; Spect=spectinomycin; Strep=streptomycin

Sequencing Primers: You cannot use the T7 universal forward primer for sequencing Duet vectors.

Name Use Sequence
Duetseq Forward, 1st MCS, pETDuet
(Binds further upstream than pET Upstream)
5'-GGTGCATGCAAGGAGATGG
DuetDOWN1 Reverse, 1st MCS, all Duet vectors 5'-GATTATGCGGCCGTGTACAA
DuetUP2 Forward, 2nd MCS, all Duet vectors 5'-TTGTACACGGCCGCATAATC
pColaUP Forward, 1st MCS, pCDFDuet, pCOLADuet, pRSFDuet
(Identical to ACYCDuetUP1)
5'-GGATCTCGACGCTCTCCCT
pET Upstream Forward, 1st MCS, pETDuet 5'-ATGCGTCCGGCGTAGA
T7Rev Reverse, 2nd MCS, all Duet vectors 5'-GCTAGTTATTGCTCAGCGGT