DNA insert from MON87460 vector PV-ZMAP595
The T-DNA insert contains the following gene cassettes: Bacillus subtillus cold shock protein (cspB) and Escherichia coli neomycin phosphotransferase II (nptII). 

Transcription of cspB is under control of the Oryza sativa actin 1 promoter and Agrobacterium tumefaciens transcript 7 gene 3' untranslated region. The transcript initially contains an O. sativa actin 1 intron for enhanced gene expression of cspB. The sequence is removed (spliced) prior to protein translation. Constitutive expression of cspB is expected due to the actin promoter.

Transcription of nptII is under control of the Cauliflower mosaic virus (CaMV) 35S promoter and A. tumefaciens nopaline synthase terminator. High levels of transcription are expected due to the CaMV promoter.

Note:
- The coding sequence of cspB has been codon optimized for optimal expression within plant cells.
- Southern blot analysis indicated that no vector backbone sequences were inserted into the parental genome
- Southern blot analysis indicated that the parental genome contains a single insertion
- Sequencing analyses confirm the Southern blot analyses.
- A 22 base pair deletion of genomic DNA at the insert-to-plant DNA junction occurred.
- loxP sites can be found in the parental genome and could potentially allow for the excision of the nptII cassette by CRE recombinase.

DNA insert from TC1507 vector PHI8999A
DNA fragment PHI8999A contains two adjacent plant gene expression cassettes for Bacillus thuringiensis cry1F and Streptomyces viridochromogenes pat.

Transcription of cry1F is directed by the promoter and first exon and intron of the maize (Zea mays) ubiquitin gene and terminates at the Agrobacterium tumefaciens ORF25 terminator.

Transcription of the pat gene commences from the Cauliflower mosaic virus (CaMV) 35S promoter and ends at the CaMV 35S terminator.

Note:
- The coding sequence of both genes has been optimized to achieve a high level of expression in maize.
- The sequences of the complete cry1F and pat are identical to those in the original plasmid.
- The CRY1F protein includes the F604K (phenylalanine to lysine at position 604) amino acid substitution, which was introduced to create a specific restriction site for cloning purposes.

DNA insert from 59122 vector PHP17662:
Transcription of Bacillus thuringiensis cry34Ab1 starts at Zea mays ubiquitin gene promoter and terminates at the Solanum tuberosum proteinase inhibitor II gene terminator. Transcription of B. thuringiensis cry35Ab1 commences from the (Triticum aestivum (wheat) peroxidase gene promoter and stops at another S. tuberosum proteinase inhibitor II gene terminator.

Note:
- The coding sequence of cry34Ab1 and cry35Ab1  has been adapted to the codon usage in maize as to achieve optimal expression in planta.
- The cry34Ab1 and cry35Ab1 were cloned from B. thuringiensis strain PS149B1.
- 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 peroxidase 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.

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