Genetic elements from PV-ZMAP1043:
Transcription of the Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4 epsps) gene commences from the Cauliflower Mosaic Virus (CaMV) 35S enhanced promoter and terminates at the A. tumefaciens nopaline synthase (nos) terminator. The cp4 epsps coding sequence is the codon optimized coding sequence of the aroA gene from A. tumefaciens strain CP4 encoding CP4 EPSPS.

Note: A single insertion event without vector background in the parental line was previously determined using DNA sequencing.

Genetic elements from PV-ZMAP595:
Two genetic cassettes were inserted into the genome:  cspB and nptII. The cspB cassette is under the control of the Rice (Oryza sativa) actin 1 promoter and contains a non-translated rice actin 1 intron, Bacillus subtilis Cold Shock Protein B (cspB), and the polyadenlytaion sequence from A. tumefaciens transcript 7. The neomycin phosphotransferase II (nphII) gene is under the control of a CaMV 35S promoter and terminates at the A. tumefaciens nos terminator. The nphII gene  is a selectable marker for use in transformation.

Note: A single insertion containing each genetic cassette was confirmed in the parental line.

Genetic elements 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 gene 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.

Genetic elements from PHI8999A:
DNA fragment PHI8999A contains two adjacent plant gene expression cassettes for the cry1F and pat genes. Transcription of the cry1F gene was directed by the promoter and first exon and intron of the maize ubiquitin gene. The 3' terminator sequence used was the 3' polyadenylation signal from ORF25 (A. tumefaciens) (see footnote*). Transcriptional regulation of the pat gene from Streptomyces viridochromogenes was via promoter and terminator sequences derived from the 35S transcript of CaMV. The coding sequence of both genes has been optimized to achieve a high level of expression in maize.

Note: The sequences of the complete cry1F and pat genes are identical to those in the original plasmid. The proteins produced in the modified plants are the ones intended, including a leucine residue (replacing a phenylalanine) at position 604 (of 605 amino acids in total) of Cry1F. This modification was introduced to create a specific restriction site for cloning purposes.

Genetic elements from PHP17662:
The cry34Ab1 and cry35Ab1 were cloned from B. thuringiensis strain PS149B1 and the coding sequence of both genes has been adapted to the codon usage in maize as to achieve expression in the maize plant. Transcription of Cry34Ab1 starts at Z. mays ubiquitin gene promoter and terminates at the Solanum tuberosum proteinase inhibitor II gene terminator. Expression of Cry35Ab1 commences from the wheat peroxidase gene promoter and stops at another S. tuberosum proteinase inhibitor II gene terminator.

Note: Sequence analysis of 59122 done by the European Food Safety Authority indicated that this LMO contains one complete copy of the T-DNA of PHP17662 without internal rearrangements. All three gene cassettes, cry34Ab1, cry35Ab1 and pat, are intact within the transgenic event. The DNA sequences of the genes in 59122 are identical to those in the original plasmid except for two nucleotide differences in the wheat peroxidise promoter. At the 5' T-DNA end a deletion of 22 bp is observed and at the 3' T-DNA end a deletion of 25 bp is observed. The absence of vector backbone in maize 59122 was also demonstrated.

Genetic elements from PV-ZMIR10871:
The DvSnf7p sequence is the partial coding sequence of the Snf7 gene from Diabrotica virgifera virgifera encoding the SNF7 subunit of the ESCRT-III complex. The DvSnf7 suppression cassette contains two 240 bp DvSnf7p sequences in an inverted orientation. There is an intervening sequence of 150 nucleotides between the two DvSnf7p sequences. When the suppression cassette is transcribed from  the CaMV 35S enhanced promoter, the RNA expressed forms a hairpin loop thereby allowing the formation of double stranded DvSnf7 RNA. Transcription terminates at the Pisum sativum rubisco small subunit The DvSnf7p sequences in the suppression cassette produce a 240 bp dsRNA that upon transcription triggers the RNAi mechanism.

The transformation also contains expression cassettes for cry3Bb1 and epsps. Expression of cry3Bb1 commences from Z. mays physical impedance induced protein promoter and continues, transcribing wheat cab, rice actin 1 intron, and B. thuringiensis cry3Bb1, and then terminates at the rice alpha tublin 1 terminator.

Note: Sequencing, PCR, and bioinformatic analysis indicated that a single, intact insert of the DvSnf7 suppression cassette and the cry3Bb1 and cp4 epsps expression cassettes were stably integrated into the maize genome. Analyses also indicated the absence of the plasmid backbone DNA in MON 87411 maize.