DNA insert from MON87427 (PV-ZMAP1043)
The parental maize contributed one genetic cassette for Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps).

Transcription cp4-epsps commences from the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and ends at the A. tumefaciens nopaline synthase (nos) gene terminator. The transcript contains a Zea mays heat shock protein 70 (hsp70) intron, Arabidopsis thaliana N-terminal chloroplast transit peptide sequence and cp4-epsps. The CaMV enhanced 35S promoter-hsp70 combination promotes gene expression in female and vegetative tissues, but not in male reproductive tissues (pollen microspores and tapetum). The chloroplast transit peptide directs the protein to accumulate in the chloroplast.

Note:
- Southern blot analyses indicate that a single copy of the T-DNA was inserted at a single site in the parental maize genome and no plasmid vector backbone sequences were detected to have been integrated. DNA sequencing analyses further indicated that the expected T-DNA sequences were integrated.
-The cp4-epsps coding sequence is the codon optimized coding sequence of the aroA gene from Agrobacterium sp. strain CP4 encoding CP4 EPSPS.

DNA insert from MON94804 (vector PV-ZMAP527892)
The parent maize contributed one genetic cassette encoding a microRNA interference suppression cassette. Transcription of the this gene cassette is under control of the Rice tungro bacilliform virus 1 promoter and a synthetic maize terminator (GST43). The initial transcript includes both a maize heat shock protein 70 intron to enhance transcription and a gibberellic acid 20 oxidase suppression sequence, comprised of two inverted repeats of gibberellic acid 20 oxidase flanked and separated by Oryza sativa precursor miRNA MIR1425 sequences. Upon transcription, the inverted sequences base pair to form a hairpin structure (pri-miRNA), which will trigger an RNA interference response in the cells, resulting in processing of the pri-miRNA into mature microRNA. Thus, no translation of this coding sequence is expected to occur. For more information, see the section "Please describe how the expression of the gene(s) was affected" below.

Note:

DNA insert from NK603 (vector PV-ZMGT32)
The parental maize contributes two adjacent genetic cassettes for Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps).

In the first (5' end) expression cassette, the cp4-epsps gene is under the transcriptional regulation of an Oryza sativa actin promoter and an A. tumefaciens nopaline synthase gene (nos) terminator. An O. sativa actin intron is also present in the transcript for enhanced expression of the coding sequence. The second cassette consists of another cp4-epsps gene regulated by a Cauliflower mosaic virus enhanced 35S promoter (containing a duplicated enhancer region) and a nos terminator. Similarly, an intron from the Zea mays heat shock protein 70 (hsp70) was included for enhancing expression of the coding sequence. Both promoters of the gene cassettes are expected to promote high levels of transcription.

Note: