DNA insert from NK603 vector 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.


DNA insert from T25 vector pDH51
The parental genome contains a Streptomyces viridochromogene phosphinothricin N-acetyltransferase gene (pat) gene cassette. Transcription of pat is under control of a Cauliflower mosaic virus 35S promoter and 35S terminator. Expression of pat is therefore expected to occur at elevated levels.

The inserted DNA additionally contains a fragment of the bacterial Escherichia coli beta-lactamase gene and the plasmid vector origin of replication (for bacterial cells). These are not anticipated to be active in plant cells.

Note:
- The pat coding sequence was codon optimized for expression in plants without resulting in amino acid changes in the translated protein.
- A single pat cassette is present in the inserted DNA.


DNA insert from DAS40278 vector pDAS1740
The LMO was generated using the Whiskers-mediated transformation method. Sphingobium herbicidovorans aryloxyalkanoate dioxygenase-1 (aad-1) is under the control of Zea mays ubiquitin gene promoter and Z. mays root preferential cationic peroxidase terminator. Elevated levels of transcription are expected to occur due to the constitutive nature of the ubiquitin promoter.

Note:
- The aad-1 coding sequence was optimized for expression in the plant.
- Southern blot analysis indicated that a single complete copy of the transformation cassette was stably integrated into the host genome at a single locus
- No integration of the vector backbone occurred.


Kindly refer to the parental LMO records for more information.