bar gene cassette (Anti-sense orientation)
The transcription of the Streptomyces hygroscopicus phosphinothricin N-acetyltransferase (bar; PAT protein) gene is under the control of the duplicated Cauliflower Mosaic Virus 35S promoter and the Glycine max vegetative storage protein terminator. To enhance translation, the Tobacco Etch Virus 5' untranslated region was included at the 5' end of the bar transcript.

Homeodomain-leucine zipper 4 gene cassette (Sense orientation)
Transcription of the Helianthus annuus Homeodomain-leucine zipper 4 (HaHB4) gene starts at the HaHB4 promoter and terminates at the Agrobacterium tumefaciens nopaline synthase gene terminator. The promoter activity is excepted to increase in response to abiotic stresses, ethylene and insect feeding, resulting in accumulations of HaHB4 transcript and HAHB4 protein.

HaHB4 nucleotide changes:
- Deletion of nucleotides 76 to 87
- A to G substitution at position 22 (codon change from AAA to AGA)
- G to A substitution at position 351 (silent point mutation - CAG to CAC)
- A to C substitution at position 401 (silent point mutation - GCA to GCC)
- T to C substitution at position 432 (codon change TTT to CTT)
- C to T substitution at position 551 (codon change CCT to CTT)

HB4 amino acid changes:
- Four amino acid deletion (residue numbers 7 to 10 - results in numbering change in residues)
- Lysine to Arginine substitution at position 22 (K22 -> R18)
- Phenylalanine to Leucine substitution at position 159 (F159 -> L155)
- Proline to Leucine substitution at position 175 (P175 -> L171)

Note:
- Southern blot, next generation sequencing and PCR analyses indicate that the plasmid backbone was absent in the genome.
- Southern blotting and sequencing confirmed the presence of a single, T-DNA insert.
- PCR analysis indicated that the transgene insert followed Mendelian inheritance.
- Bioinformatic analysis indicated that no new open read frames were created as a result of the T-DNA insertion.