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Iterative algorithm-guided design of massive strain libraries, applied to itaconic acid production in yeast.

Young EM, Zhao Z, Gielesen BEM, Wu L, Benjamin Gordon D, Roubos JA, Voigt CA
Metab Eng. 2018 Jul;48:33-43. doi: 10.1016/j.ymben.2018.05.002. Epub 2018 May 9. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
127710pEMY07AB-PROLevel 0 destination vector for a promoter with A - B scars. Clone in with BpiI
127711pEMY07AB-ORFLevel 0 destination vector for a coding sequence with B - C scars. Clone in with BpiI
127712pEMY07CD-TERLevel 0 destination vector for a terminator with C - D scars. Clone in with BpiI.
127713pY128Level 1 yeast shuttle vector for a transcription unit with A - D scars. Clone in with BsaI. Has a high-copy yeast replicon and KanMX selection.
127714pEMY01AB-PROIntegrative yeast promoter characterization plasmid. Clone in with BpiI. Has ChrXV homology upstream and an mVenus fragment downstream.
127716pDSM-5aLevel 1 destination vector for a transcription unit, pathway position 1. Clone a transcription unit in with BsaI.
127717pDSM-abLevel 1 destination vector for a transcription unit, pathway position 2. Clone a transcription unit in with BsaI.
127718pDSM-bcLevel 1 destination vector for a transcription unit, pathway position 3. Clone a transcription unit in with BsaI.
127719pDSM-cdLevel 1 destination vector for a transcription unit, pathway position 4. Clone a transcription unit in with BsaI.
127720pDSM-deLevel 1 destination vector for a transcription unit, pathway position 5. Clone a transcription unit in with BsaI.
127721pDSM-efLevel 1 destination vector for a transcription unit, pathway position 6. Clone a transcription unit in with BsaI.
127722pDSM-f3Level 1 destination vector for a transcription unit, pathway position 7. Clone a transcription unit in with BsaI.
127723pDSM-5a-Pact1-ITE-Tadh1Yeast pathway position 1. ITE transcription unit with the ACT1 promoter and ADH1 terminator.
127724pDSM-5a-Ppre3-ITE-Teno1Yeast pathway position 1. ITE transcription unit with the PRE3 promoter and ENO1 terminator.
127725pDSM-5a-Pact1-ITE-Tdpp1Yeast pathway position 1. ITE transcription unit with the ACT1 promoter and DPP1 terminator.
127726pDSM-5a-Phta1-ITE-Tvma2Yeast pathway position 1. ITE transcription unit with the HTA1 promoter and VMA2 terminator.
127727pDSM-bc-Ptdh3-CAD-Ttdh1Yeast pathway position 3. CAD transcription unit with the TDH3 promoter and TDH1 terminator.
127728pDSM-bc-Prps3-CAD-Tcyc1Yeast pathway position 3. CAD transcription unit with the RPS3 promoter and CYC1 terminator.
127729pDSM-bc-Peft2-CAD-Tklura3Yeast pathway position 3. CAD transcription unit with the EFT2 promoter and K. lactis URA3 terminator.
127730pDSM-bc-Psmtef1-CAD-Tyhi9Yeast pathway position 3. CAD transcription unit with the S. mikatae TEF1 promoter and YHI9 terminator.
127731pDSM-cd-Pfba1p-ACO-Tgpm1Yeast pathway position 4. ACO transcription unit with the FBA1 promoter and GPM1 terminator.
127732pDSM-cd-Pykt6-ACO-Tprm9Yeast pathway position 4. ACO transcription unit with the YKT6 promoter and PRM9 terminator.
127733pDSM-cd-Ptef1-ACO-Tyol036wYeast pathway position 4. ACO transcription unit with the TEF1 promoter and YOL036w terminator.
127734pDSM-cd-Psptdh3-ACO-Tagtef1Yeast pathway position 4. ACO transcription unit with the S. paradoxus TDH3 promoter and A. gossypii TEF1 terminator.
127735pDSM-de-Ppgk1-PYC-Ttpi1Yeast pathway position 5. PYC transcription unit with the PGK1 promoter and TPI1 terminator.
127736pDSM-de-Ppxr1-PYC-Tnat1Yeast pathway position 5. PYC transcription unit with the PXR1 promoter and NAT1 terminator.
127737pDSM-de-Phhf2-PYC-Tadh1Yeast pathway position 5. PYC transcription unit with the HHF2 promoter and ADH1 terminator.
127738pDSM-de-Pfba1-PYC-Trpl15aYeast pathway position 5. PYC transcription unit with the FBA1 promoter and RPL15A terminator.
127739pDSM-ef-Ptef1-CSC-Tpdc1Yeast pathway position 6. CSC transcription unit with the TEF1 promoter and PDC1 terminator.
127740pDSM-ef-Pzuo1-CSC-Tvma16Yeast pathway position 6. CSC transcription unit with the ZUO1 promoter and VMA16 terminator.
127741pDSM-ef-Ppfy1-CSC-Taip1Yeast pathway position 6. CSC transcription unit with the PFY1 promoter and AIP1 terminator.
127742pDSM-ef-Psbtdh3-CSC-Tefm1Yeast pathway position 6. CSC transcription unit with the S. bayanus TDH3 promoter and EFM1 terminator.
127743pDSM-f3-Peno2-ACDH-Ttal1Yeast pathway position 7. ACDH transcription unit with the ENO2 promoter and TAL1 terminator.
127744pDSM-f3-Pvma6-ACDH-Trps14aYeast pathway position 7. ACDH transcription unit with the VMA6 promoter and RPS14A terminator.
127745pDSM-f3-Prpl28-ACDH-Tecm10Yeast pathway position 7. ACDH transcription unit with the RPL28 promoter and ECM10 terminator.
127746pDSM-f3-Ptdh3-ACDH-Trpl3Yeast pathway position 7. ACDH transcription unit with the TDH3 promoter and RPL3 terminator.
127747pDSM-ab-Psptef1-NAT-Ttip1Yeast pathway position 2. NatMX transcription unit with the S. paradoxus TEF1 promoter and TIP1 terminator. Nourseothricin resistance.
127751pEMY01CD-TERIntegrative yeast terminator characterization plasmid. Clone in with BpiI. Has mVenus fragment upstream and a KanMX fragment downstream.

Antibodies from Article