Development of a dedicated Golden Gate Assembly Platform (RtGGA) for Rhodotorula toruloides.
Bonturi N, Pinheiro MJ, de Oliveira PM, Rusadze E, Eichinger T, Liudziute G, De Biaggi JS, Brauer A, Remm M, Miranda EA, Ledesma-Amaro R, Lahtvee PJ
Metab Eng Commun. 2022 May 23;15:e00200. doi: 10.1016/j.mec.2022.e00200. eCollection 2022 Dec.
(Link opens in a new window)
PubMed
(Link opens in a new window)
Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 187626 | (P) insUP_ku70 (SBE138) (B) | the insertional region upstream for the deletion of the KU70 gene in R. toruloides. Overhangs P/B |
| 187627 | (S) insD_ku70 (Q) (SBE139) | the insertional region downstream for the deletion of the KU70 gene in R. toruloides. Overhangs S/Q |
| 187628 | (F) pADH2 (G)_P2 (SBE113) | promoter of the gene alcohol dehydrogenase 2 from R. toruloides fro the P2 position (F/G) |
| 187629 | (I) pADH2 (J) P3 (SBE114) | promoter of the gene alcohol dehydrogenase 2 from R. toruloides for the P3 position (I/J) |
| 194990 | (I) pGPD (J)_P3 (SBE111) | promoter of the gene glyceraldehyde-3-phosphate dehydrogenase from R. toruloides for a P3 position (I/J) |
| 194991 | (I) pXYL (J) P3 (SBE108) | promoter of the gene xylose reductase from R. toruloides for a position P3 |
| 194992 | (F)pGPD(G)_P2 (SBE110) | promoter of the gene glyceraldehyde-3-phosphate dehydrogenase from R. toruloides for a P2 position (overhangs F/G) |
| 194993 | (R)pGPD1(D)_P1(SBE109) | promoter of the gene glyceraldehyde-3-phosphate dehydrogenase from R. toruloides for a P1 position (overhangs R/D) |
| 194994 | (D) crtE (E)_G1 (SBE115) | Rhodotorula toruloides gene geranylgeranyl diphosphate synthase (crtE, RHTO_02504) for G1 |
| 194995 | (G) crtI (H) G2 (SBE116) | Rhodotorula toruloides gene phytoene dehydrogenase (crtI, RHTO_04602) for G2 |
| 194996 | (J) crtYB(K) G3 (SBE117) | Rhodotorula toruloides gene phytoene synthase/lycopene cyclase (crtYB, RHTO_04605) for G3 |
| 194997 | (E) tNOS (F) _T1 (SBE118) | terminator from Agrobacterium tumefaciens nopaline synthase (tNOS, MF116010) for T1 position (E/F) |
| 195006 | (H) tNOS (I) _T2 (SBE119) | terminator from Agrobacterium tumefaciens nopaline synthase (tNOS, MF116010) for T2 position (H/I) |
| 195010 | (K) tNOS (S)_T3 (SBE120) | terminator from Agrobacterium tumefaciens nopaline synthase (tNOS, MF116010) for T3 position (K/S) |
| 195011 | (B) G418 marker (R) (SBE 137) | geneticin (G418) resistance genes codon-optimized for R. toruloides and expressed under pXYL and tNOS |
| 195012 | (B) BLE marker(R) (SBE262) | bleomycin (BLE) resistance genes codon-optimized for R. toruloides and expressed under pXYL and tGPD |
| 195013 | (B) HYG marker(R) (SBE264) | hygromycin (HYG) resistance genes codon-optimized for R. toruloides and expressed under pXYL and tGPD |
| 195014 | (B) NAT marker (R) (SBE173) | nourseothricin (NAT) resistance genes codon-optimized for R. toruloides and expressed under pXYL and tNOS |
| 195015 | (R)pADH2(D)_P1(SBE214) | promoter of the gene alcohol dehydrogenase 2 from R. toruloides for the position P1 |
| 195016 | (R) pXYL (D) P1 (SBE215) | promoter of the gene xylose reductase from R. toruloides for the position P1 (R/D) |
| 195017 | (F) pXYL (G) P1 (SBE216) | promoter of the gene xylose reductase from R. toruloides for the position P2 (F/G) |
| 195046 | ku70 insUP+ pXYL-G418-tNOS-pXYL-crtE-tNOS-pGPD-crtI-tNOS-pADH2-crtYB-tNOS+ku70 insD (SBE144) | cassette for the deletion of ku70 and overexpression of crtE, crtI, and crtYB under different combination of promoters |
| 195047 | ku70 insUP+ pXYL-G418-tNOS-pADH2-crtE-tNOS-pGPD-crtI-tNOS-pXYL-crtYB-tNOS+ku70 insD (SBE145) | cassette for the deletion of ku70 and overexpression of crtE, crtI, and crtYB under different combination of promoters |
| 195048 | ku70 insUP+ pXYL-G418-tNOS-pADH2-crtE-tNOS-pXYL-crtI-tNOS-pGPD-crtYB-tNOS+ku70 insD (SBE146) | cassette for the deletion of ku70 and overexpression of crtE, crtI, and crtYB under different combination of promoters |
| 195049 | ku70 insUP+pPGI+NAT+tNOS+ku70 insD (SBE217) | integration cassette for ku70 deletion and expression of NAT gene under pPGI for characterization |
| 195050 | ku70 insUP+pADH2+NAT+tNOS+ku70 insD (SBE174) | integration cassette for ku70 deletion and expression of NAT gene under pADH2 for characterization |
| 195051 | ku70 insUP+pGPD+NAT+tNOS+ku70 insD (SBE175) | integration cassette for ku70 deletion and expression of NAT gene under pGPD for characterization |
| 195052 | ku70 insUP+pXYL+NAT+tNOS+ku70 insD (SBE178) | integration cassette for ku70 deletion and expression of NAT gene under pXYL for characterization |
| 195053 | ku70 insUP+pFBA+NAT+tNOS+ku70 insD (SBE179) | integration cassette for ku70 deletion and expression of NAT gene under pFBA for characterization |
| 195054 | ku70 insUP+pLDP1+NAT+tNOS+ku70 insD (SBE193) | integration cassette for ku70 deletion and expression of NAT gene under pLDP1 for characterization |