DNA insert from PV-ZMAP1043
Transcription of  Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4 epsps) commences from the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and terminates at the A. tumefaciens nopaline synthase (nos) terminator. The transcript contains a Zea mays heat shock protein 70 (hsp70) intron, an Arabidopsis thaliana N-terminal chloroplast transit peptide sequence for chloroplast targeting of the protein 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).

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 PV-ZMIR245
Two insecticidal protein expression cassettes were inserted into the genome. Bacillus thuringiensis cry1A.105 expression is under the control of the CaMV 35S enhanced promoter, which first transcribes wheat (Triticum aestivum) 5' untranslated region of the chlorophyll a/b-binding protein (cab) and a rice actin 1 intron before transcribing cry1A.105. Transcription terminates at the wheat heat shock protein 17.3 terminator. Expression of the B. thuringiensis cry2Ab2 starts at the Figwort mosaic virus (FMV) promoter, which transcribes the Zea mays heat shock protein 70 (hsp70), then the Z. mays transit peptide and the cry2Ab2 coding sequence, before terminating at the nos terminator.

Note:
- The Cry2Ab2 coding sequence was modified for optimal expression in plants.
- South blot analysis confirmed that single insertions of both cry2Ab2 and cry1A.105, as well as no vector backbone were present and in the parent.
- A deletion removed the duplicated enhancer elements compared to the original CaMV e35S promoter in PV-ZMIR245.
- The selectable marker, nphII, cassette was bred out of the parental line and became not associated with this transformation event.


DNA insert from PV-ZMHT507801
The Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat) gene is under the control of the Andropogon gerardii ubiquitin  promoter and the Oryza sativa alpha-amylase/trypsin inhibitor terminator. The transcript includes the A. gerardii 5' untranslated leader sequence and an intron from ubiquitin before (5') coding sequence of pat.

The Stenotrophomonas maltophilia dicamba monooxygenase (dmo) gene is under control of the Peanut chlorotic streak caulimovirus (PC1SV) full-length transcript promoter and the Triticum aestivum (wheat) heat shock protein 17 terminator. The transcript produced contains a wheat chlorophyll a/b-binding 5' untranslated leader sequence (for improved gene expression), an O. sativa actin 1 untranslated region and intron (for improved gene expression), the untranslated  and targeting region of Petunia hybrida chloroplast transit peptide 4 (for chloroplast targeting of the protein) and dmo.

Note:
- Originally, the plasmid vector contained two T-DNA elements that were inserted during the initial transformation event: one containing the dmo and pat expression cassettes, and a second containing an Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) expression cassette. The cp4-epsps expression cassette is regulated by the O sativa actin 1 promoter and 5′ untranslated leader, O. sativa intron, the Arabidopsis thaliana chloroplast targeting peptide 2 targeting sequence, and the A. tumefaciens nopaline synthase  3′ untranslated region. Subsequent traditional breeding, segregation, selection, and screening were used to isolate those plants that contain the dmo and pat expression cassettes (T-DNA I) and do not contain the cp4-epsps expression cassette (T-DNA II).
- Molecular characterization of MON87419 indicated that a single copy of T-DNA I was integrated into the maize genome at a single intact locus that includes all expected elements within the insert, with the exception of incomplete Right and Left Border sequences. These analyses also showed no PV-ZMHT507801 backbone elements or T-DNA II sequences were present in the event.


DNA insert from PV-ZMGT32
The plant expression plasmid vector, PV-ZMGT32 contains two adjacent plant gene expression cassettes each containing a single copy of the 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 a 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 an CaMV enhanced 35S promoter (containing a duplicated enhancer region) and a nos terminator. Similarly, an intron from the maize heat shock protein 70 (hsp70) was included for enhancing expression of the coding sequence. Both promoters of the gene cassettes are expected to promoter high levels of transcription.

Note:
- The parental NK603 line contained a single, intact insertion containing both cp4 epsps gene cassettes.
- Due to restriction digest prior to particle bombardment, the vector backbone, containing E. coli neomycin phosphotransferase II and origin of replication, were not incorporated into the parental genome.


For more information, kindly refer to the parental line records.