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pRec445

3'Nos. NPT II. Actin intron. Ds (3'Ac ). Actin promoter. CsVMV promoter. Ds (5'Ac ). TATA box. +1. LB. Actin intron. Gnl1-3 cDNA. 3' GS-1b. Spectinomycin. cos site. 3'Nos. tetR. RB. ColE1. tetA. pRec445. Rep operon. 47167 bp. virB. from pTiBo542. oriT. virG operon.

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pRec445

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  1. 3'Nos NPT II Actin intron Ds (3'Ac ) Actin promoter CsVMV promoter Ds (5'Ac ) TATA box +1 LB Actin intron Gnl1-3 cDNA 3' GS-1b Spectinomycin cos site 3'Nos tetR RB ColE1 tetA pRec445 Rep operon 47167 bp virB from pTiBo542 oriT virG operon virC operon Ctl oriV Supplementary file 2. Map of the pREc 445 Plasmid Containing the Gln1-3 cDNA Under the Control of the CsVMV Promoter. RB and LB represent the right and left borders of the T-DNA. Ds(3’Ac) and Ds (5’Ac) represent the Ds transposable element used to further remove the selection marker conferring resistance to kanamycin. NptII is the neomycin phosphotransferase gene with the Actin promoter and the Actin intron and nopaline synthetase terminator (3’Nos) conferring kanamycin resistance. An Actin intron was placed between the maize Gln1-3 cDNA and the CsVMV promoter. The LB and RB extra borders, the 3’Nos, the Gln1-3 and the intron actin were used as probes to determine the pattern of the construct insertion events in the transgenic maize lines.

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