Information on the inserted DNA sequences
The transforming plasmid PV-GHGT35 carried a transfer DNA sequence comprising of two codon-optimised Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (epsps) cassettes:

(1) the first epsps coding sequence under the regulation of a chimeric transcriptional promoter (Figwort mosaic virus 35S promoter enhancer and Arabidopsis thaliana elongation factor EF-1 alpha (tsf1) promoter), tsf1 leader and intron sequences, an A. thaliana chloroplast transit peptide 2 sequence and a Pisum sativum ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) E9 transcript termination and polyadenylation sequence (T-E9).

(2) the second epsps coding sequence regulated by another chimeric transcriptional promoter (Cauliflower mosaic virus 35S enhancer and A. thaliana actin 8 (act8) promoter), act8 leader and intron sequences, A. thaliana chloroplast targeting peptide 2 and T-E9.

High levels of transcription are expected from both cassettes due to the presence of viral enhancer sequences. The EPSPS protein is expected to accumulate in the chloroplast due to the transit signal peptide.

Vector information
Monsanto constructed the double border, binary plasmid vector PV-GHGT35 for the transformation of cotton variety Coker 312. The plasmid contains a single copy of two epsps expression cassettes within the T-DNA region. The T-DNA region of PV-GHGT35 was incorporated into the target cotton genome using Agrobacterium-mediated transformation. Plasmid PV-GHGT35 also contains several genes from the plasmid backbone necessary for maintenance and selection of the plasmid that are not ultimately incorporated into the plant genome. Plasmid PV-GHGT35 contains both vegetative and bacterial origins of replication that allow replication of the plasmid in both A. tumefaciens and Escherichia coli. The plasmid contains the aad gene encoding the Tn7 adenyltransferase that provides resistance to spectinomycin and streptomycin. The plasmid also contains a sequence, known as rop, which represses the formation of RNA primer thereby allowing maintenance and copy number control of the plasmid in Escherichia coli.

Note on genetic element sizes:
The promoter for both genetic constructs are chimeric promoters containing viral enhancer sequences. Thus, the size of the promoters (Elongation factor 1 alpha and Actin 8) in the 'Genetic elements construct' reflects the size of chimeric promoters (FMV 35S enhancer + Elongation factor 1 alpha promoter; CaMV 35S enhancer + Actin 8 promoter).