Abstract: The invention relates to a process for determining the functionality of an artificial genetic element. An artificial genetic element designed to serve the same biological function as a native genetic element is introduced into cells, and the cells are subjected to conditions of high frequency transposon mutagenesis. Subsequently, a set of DNA sequences representative of a site of insertion of the transposable element is obtained from the cells and the frequency of insertion of the transposable element into the native genetic element and the artificial genetic element is determined. Comparing the frequency of transposon insertion allows for assigning a likelihood of functionality to said artificial functional element, which is high if the frequency of transposon insertion is essentially equal for both elements, and which is low if the frequency of transposon insertion is higher into the artificial genetic element.

Abstract: The invention relates to a process for assembling DNA parts into multi-kilo base long synthetic DNA constructs. The process generates multiple, synonymous DNA parts in parallel and selects in a combinatorial assembly approach for those sequence variants with the best synthesis and assembly feasibility. DNA parts are sequence optimized and partitioned into synonymous variant designs that serve as redundant building units for higher order DNA assembly. The major stages of the process are: computational partitioning and synonymous recoding of the DNA design, DNA synthesis of sequence variants pools, serial PGR to isolate sets of DNA parts and higher order assembly. As the higher-order assembly does no longer depends on successful synthesis of each DNA part, large-scale DNA designs can be quickly completed allowing for cost-effective and highly parallelised assembly of synthetic bio-designs.

Abstract: The invention relates to a process for determining the functionality of an artificial genetic element. An artificial genetic element designed to serve the same biological function as a native genetic element is introduced into cells, and the cells are subjected to conditions of high frequency transposon mutagenesis. Subsequently, a set of DNA sequences representative of a site of insertion of the transposable element is obtained from the cells and the frequency of insertion of the transposable element into the native genetic element and the artificial genetic element is determined. Comparing the frequency of transposon insertion allows for assigning a likelihood of functionality to said artificial functional element, which is high if the frequency of transposon insertion is essentially equal for both elements, and which is low if the frequency of transposon insertion is higher into the artificial genetic element.

Abstract: Compositions and methods are provided for the rapid and highly accurate identification of the entire essential genome of any organism under a given selection condition at a resolution of a few base pairs. An engineered transposon bearing an adapter sequence for ultra high throughput adaptor-based sequencing is employed for hyper-saturated transposon mutagenesis. Transposon junctions are subsequently isolated and collectively amplified through a shared parallel PCR strategy such that a second adaptor sequence is further incorporated into template DNA so that the first adaptor sequence and the second adaptor sequence flank the 5? and 3? regions of the sample DNA, respectively. Sample DNA is then sequenced in an ultra high-throughput adaptor-based DNA sequencer using adaptor primers. Transposon insertion sites are mapped onto the organism's genome, allowing for the algorithmic identification of essential genetic elements based on genomic transposition frequency.

Abstract: Compositions and methods are provided for the rapid and highly accurate identification of the entire essential genome of any organism under a given selection condition at a resolution of a few base pairs. An engineered transposon bearing an adapter sequence for ultra high throughput adaptor-based sequencing is employed for hyper-saturated transposon mutagenesis. Transposon junctions are subsequently isolated and collectively amplified through a shared parallel PCR strategy such that a second adaptor sequence is further incorporated into template DNA so that the first adaptor sequence and the second adaptor sequence flank the 5? and 3? regions of the sample DNA, respectively. Sample DNA is then sequenced in an ultra high-throughput adaptor-based DNA sequencer using adaptor primers. Transposon insertion sites are mapped onto the organism's genome, allowing for the algorithmic identification of essential genetic elements based on genomic transposition frequency.