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Table 1 Plasmids used in this study

From: A synthetic system links FeFe-hydrogenases to essential E. coli sulfur metabolism

Name Constructs Backbone Resistance Source
Hydrogenase activity in vivo A
pET.mp1 ca HydE ca HydA pETDuet-1 Ampicillin Matthew Posewitz[31]
pCDF.mp2 ca HydF ca HydG pCDFDuet-1 Spectinomycin Matthew Posewitz[31]
pACYC.ew3 da PFOR   pACYCDuet-1 Chloramphenicol This work
pACYC.ew4 da PFOR so FD pACYCDuet-1 Chloramphenicol This work
pACYC.ew5 da PFOR zm FD pACYCDuet-1 Chloramphenicol This work
pACYC.ew6 da PFOR cr FD pACYCDuet-1 Chloramphenicol This work
pACYC.ew7 da PFOR ca FD pACYCDuet-1 Chloramphenicol This work
pACYC.ew8 so FD   pACYCDuet-1 Chloramphenicol This work
pACYC.ew9 zm FD   pACYCDuet-1 Chloramphenicol This work
pACYC.ew10 cr FD   pACYCDuet-1 Chloramphenicol This work
pACYC.ew11 ca FD   pACYCDuet-1 Chloramphenicol This work
FNR-supported growth B
pCDF.ew12 zm FNR   pCDFDuet-1 Spectinomycin This work
pACYC.ew13 so FD zm SIR pACYCDuet-1 Chloramphenicol This work
pACYC.ew14 zm FD zm SIR pACYCDuet-1 Chloramphenicol This work
pACYC.ew15 cr FD zm SIR pACYCDuet-1 Chloramphenicol This work
pACYC.ew16 ca FD zm SIR pACYCDuet-1 Chloramphenicol This work
pACYC.ew17 zm SIR   pACYCDuet-1 Chloramphenicol This work
Hydrogenase-supported growth and selection C
pACYC.ew18 cr HydEF cr HydG pACYCDuet-1 Chloramphenicol This work
pET.ew19 so FD zm SIR pETDuet-1 Ampicillin This work
pCDF.ew20 cr HydA   pCDFDuet-1 Spectinomycin This work
pCDF.ew21 ca HydA   pCDFDuet-1 Spectinomycin This work
pCDF.ew22 cs HydA   pCDFDuet-1 Spectinomycin This work
  1. Duet vector backbones (Novagen) were used for all protein expression. Complete vector sequences are provided in additional file 1. A) Plasmids mp1-ew11 were used to generate PFOR-driven hydrogenase activity data for Figure 4. B) Plasmids ew12-ew17 were used to assess knockout strain insulation for Figure 2. They were also used to measure ferredoxin performance in Figure 4. C) Plasmids ew18-ew22 facilitated the hydrogenase O2 tolerance measurements in Figure 3. They were also used in the hydrogenase-supported growth curves in Figure 5 and for the genetic selection.