DNA insert from DP202216 (vector PHP40099)
The parental maize contains two gene cassettes: Zea mays MADS-box transcription factor zmm28 and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat).
-Transcription of zmm28 is under control of the Z. mays gos2 promoter and the Solanum tuberosum proteinase inhibitor II gene (pinII) terminator. In addition to the zmm28 coding sequence, a Z. mays ubiquitin 1 intron is present at the 5' end of the transcript to enhance gene expression. The gos2 promoter is expected to promote constitutive expression in all plant tissues.
-Transcription of pat is under control of the Z. mays ubiquitin promoter and the pinII terminator. In addition to the pat coding sequence, the Z. mays ubiquitin 1 5' untranslated region and intron were added to the 5' end to enhance gene expression. The ubiquitin promoter is expected to promote high levels of constitutive expression in all plant tissues.

Note:
- The sequence of zmm28 is comprised of the 5’ UTR at bp 2,749-2,808 (60 bp long), coding sequence at bp 2,809-3,564 (756 bp long) and the 3’ UTR at bp 3,565-3,605 (41 bp long).
- The pat coding sequence was optimized for expression in maize.
- Southern-by-Sequencing analysis indicted that the DP202216 line contained a single T-DNA insertion containing the two gene cassettes without any rearrangements or integration of the vector backbone.

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: