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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

1 2 3 4 5 6 7 81 2 3 4 5 6

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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

1 2 3 4 5 6 7 81 2 3 4 5 6

1 2 3 4 5 6 7 1 2 3 4 5 6

1 2 3 4 5 6 7

CHI pet. in pMY4

Lanes 1- 4: clones 1,2,5,6 of CHI in pmy4 uncut;

Lane5: marker; Lane6: pmy4 uncut

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

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Restricted pMY4 Not1, Sal1

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Cloning of 4CL into pRS305 and CUP1_CYC1 cassette into pRS306

1 2 3 4 5 6 7

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: MarkerChecking of the ligation by plasmids size:

PCU416 5300 bp

PRS306+CASSETTE 5031bp

4CL PMY5 8213bp

4CL PRS305(+CASSETTE) 7707bp

1 2 3 4 5 6 7 8

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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

Lane1: CHI_prs304_sac1_kpn; Lane 2: marker

Lane 1: PRS303_CUP1 SacI, ApaI; Lane 2: PRS313_CUP1

SacI, ApaI ; Lane 3: MCS _PFA6 IN PMY7 NotI, ClaI; Lane 4:

marker

1 2 1 2 3 4 5 6 7 8 9 10

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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

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

1 2 3 4 5 6 7 8

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Lanes 1-2: CHI PRS304 clones 5,6,7; Lanes 4-6:

CHS PRS306 clones 5,6,7; Lane 7: PMY4;

Lane8: PCU416

Lane1: CHI PRS304 Not1, Sal1; Lane 2: CHI PMY4 Not1, Sal1; Lane3: CHS PRS306 EcoRI, Xho; Lane4: CHS PCU416 EcoRI, Xho; Lane 5: marker