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Table 1 Overview of constructs used to engineer ent-kaurene production in R. toruloides

From: Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides

PlasmidsGenotypes/featuresABFPUB registry ID
Construct 1PANT-GfKS-HYGRABFPUB_1
Construct 2PGAPDH-GfKS-NATRABFPUB_3
Construct 3PTEF1-RtERG20(F91C)-KanMXRABFPUB_6
Construct 4PTEF1-TcGGPPS(S239C,G29D)-KanMXRABFPUB_7
Construct 5PTEF1-GgFPS(F112A)-KanMXRABFPUB_8
Construct 6PTEF1-RtBTS1-KanMXRABFPUB_9
Construct 7PANT-GfKS-PTEF1-RtERG20(F91C)-NATRABFPUB_10
Construct 8PANT-GfKS-PTEF1-TcGGPPS(S239C, G295D)-NATRABFPUB_11
Construct 9PANT-GfKS-PTEF1-GgFPS(F112A)-NATRABFPUB_12
Construct 10PANT-GfKS-PTEF1-RtBTS1-NATRABFPUB_13
StrainsIFO0880 (WT)ABFPUB_14
IFO0880/PGAPDH-GfKS-NATRABFPUB_15–17
IFO0880/PANT-GfKS-HYGRABFPUB_18, 30, 34
IFO0880/PGAPDH-GfKS-NATR/PTEF1-RtERG20(F91C)-KanMXRABFPUB_47–49
IFO0880/PGAPDH-GfKS-NATR/PTEF1-TcGGPPS(S239C,G29D)-KanMXRABFPUB_19, 20, 31
IFO0880/PGAPDH-GfKS-NATR/PTEF1-GgFPS(F112A)-KanMXRABFPUB_32, 33, 35
IFO0880/PGAPDH-GfKS-NATR/PTEF1-RtBTS1-KanMXRABFPUB_38–40
IFO0880/PANT-GfKS-PTEF1-RtERG20(F91C)-NATRABFPUB_41–43
IFO0880/PANT-GfKS-PTEF1-TcGGPPS(S239C,G295D)-NATRABFPUB_21, 22, 29
IFO0880/PANT-GfKS-PTEF1-GgFPS(F112A)-NATRABFPUB_23, 26, 36
IFO0880/PANT-GfKS-PTEF1-RtBTS1-NATRABFPUB_44–46
  1. Strains and plasmids used in this study are available upon request through the Agile BioFoundry Strain Registry (http://public-registry.agilebiofoundry.org// [37])
  2. GAPDH glyceraldehyde 3-phosphate dehydrogenase, ANT, adenine nucleotide translocase, TEF1 translational elongation factor, GfKS, Gibberella fujikuroi kaurene synthase (NCBI Accession Number, Q9UVY5), RtERG20(F91C) mutant farnesyl pyrophosphate synthase from R. toruloides (PRQ75922.1), TcGGPPS(S239C,G295D) mutant geranylgeranyl pyrophosphate synthase from Taxus canadensis (AAD16018), GgFPS(F112A) mutant farnesyl pyrophosphate synthase from Gallus gallus (P08836.2) and RtBTS1 geranylgeranyl pyrophosphate synthase BTS1 from R. toruloides (PRQ72579)