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1 2 3 4 5 6. 1 2 3 4 5 6 7 8. 10000 8000 6000 5000 4000 3500 3000 2500 2000 1500 1200 1031 900 800 700 600 500. CHS tom. PCU416 . Lane1-6: clones 1-6 uncut; Lane7 : PCU416 uncut; Lane8 : marker.

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  1. 1 2 3 4 5 6 1 2 3 4 5 6 7 8 10000 8000 6000 5000 4000 3500 3000 2500 2000 1500 1200 1031 900 800 700 600 500 CHS tom. PCU416. Lane1-6: clones 1-6 uncut; Lane7: PCU416 uncut; Lane8: marker CHS TOM. PCU416 CLONE1 . Lane1: clone1 uncut; Lane2: clone1 cut with EcoRI; Lane3: clone1 cut with EcoRI, XhoI; Lane4: PCU416 uncut; Lane5: PCU416 cut EcoRI, Xho1; Lane6: marker

  2. 1 2 3 4 5 6 1 2 3 4 5 6 7 8 10000 8000 6000 5000 4000 3500 3000 2500 2000 1500 1200 1031 900 800 700 600 500 4CL tom. PMY5. Lane1-6: clones 1-6 uncut; Lane7: PMY5 uncut; Lane8: marker 4CL TOM. PMY5 CLONE 6 . Lane1: clone6 uncut; Lane2: clone6 cut with PstI; Lane3: clone6 cut with Pst, XhoI; Lane4: PMY5 uncut; Lane5: PMY5 cut Pst, Xho1; Lane6: marker

  3. 1 2 3 4 5 6 1 2 3 4 5 6 7 1 2 3 4 5 6 7 CHI pet. in pMY4 10000 8000 6000 5000 3000 2000 1031 500 Lanes 1- 4: clones 1,2,5,6 of CHI in pmy4 uncut; Lane5: marker; Lane6: pmy4 uncut Restricted pMY4 Not1, Sal1 10000 8000 6000 5000 3000 2500 2000 1500 1000 750 500 250 Lanes 1-2: clone1,clone6 CHI in pmy4 cut with Not1 and Sal1; Lane3: pmy4 cut with Not1 and Sal1; Lane4: marker; Lanes 5-6: clones 1,6 CHI in pmy4 uncut; Lane7: pmy4 uncut

  4. 1 2 3 4 5 6 7 1 2 3 4 5 6 7 8 10000 8000 6000 5000 4000 3500 3000 2500 2000 1500 1200 1031 500 Cloning of 4CL into pRS305 and CUP1_CYC1 cassette into pRS306 Lanes 1-3: clones 1,2,3 of CUP1_CYC1 cassette into pRS306; Lanes 4-6: clones 1,2,3 of 4CL into pRS305 ; Lane7: marker Lane1: CUP1_CYC1 cassette into pRS306 cut with Sac1; Lane2: PCU416 cut with Sac1; Lane3: 4CL in prs305 cut with Sac1; 4CL pmy5 cut with Sac1; Lanes 5-7: PRS306 + cassette,pcu416,4CL PRS305 uncut; Lane 8: Marker Checking of the ligation by plasmids size: PCU416 5300 bp PRS306+CASSETTE 5031bp 4CL PMY5 8213bp 4CL PRS305(+CASSETTE) 7707bp

  5. PRS305 cut with Sac1,Apa1; PRS306 cut with Sac2, Kpn1 1 2 3 4 5 6 7 Lanes 1-3: PRS305 cut with Sac1,Apa1; Lane 4: marker; Lanes 5-7: PRS306 cut with Sac2, Kpn1

  6. 1 2 3 4 5 6 7 8 9 10 1 2 1 2 3 4 Lane1: CHI_prs304_sac1_kpn; Lane 2: marker Lanes 1-3: PRS303_CUP1 clones 1,2,3; Lanes 4-6: PRS313_CUP1 clones 1,2,3; Lanes 7-9: MCS pFA6_PMY7 clones 1,2,3; Lane 10: marker Lane 1: PRS303_CUP1 SacI, ApaI; Lane 2: PRS313_CUP1 SacI, ApaI ; Lane 3: MCS _PFA6 IN PMY7 NotI, ClaI; Lane 4: marker

  7. 1 2 3 4 5 6 7 8 Lane1: PRS306 +CASSETTE Sac1, Kpn1; Lane2: PCU416 Sac1, Kpn1 ; Lane3: 4CL PRS305 Sac1, Apa1; Lane4: 4CL PMY5 Sac1, Apa1; Lane5: PRS306 uncut ;Lane6: PCU416 uncut; Lane7:4CLPRS305uncut; Lane8: MARKER

  8. 1 2 3 4 5 6 7 8 Lanes 1-2: CHI PRS304 clones 5,6,7; Lanes 4-6: CHS PRS306 clones 5,6,7; Lane 7: PMY4; Lane8: PCU416 1 2 3 4 5 Lane1: CHI PRS304 Not1, Sal1; Lane 2: CHI PMY4 Not1, Sal1; Lane3: CHS PRS306 EcoRI, Xho; Lane4: CHS PCU416 EcoRI, Xho; Lane 5: marker

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