A modular assembly cloning technique (aided by the BIOF software tool) for seamless and error-free assembly of long DNA fragments

Table 2 Sub-fragments of the Chinese hamster elongation factor 1 alpha downstream flanking region

Sub-fragment number	Start*	End*	Length	5′-flanking RE	3′-flanking RE	Clones sequenced **	Clones with the correct sequence, %
1	14545	15157	613	(Nhe I)	Xba I	4	75
2	15154	15908	755	Nhe I	Avr II	3	100
3	15905	16644	740	Nhe I	Avr II	3	100
4	16639	17327	689	Avr II	Sac I	3	100
5	17322	18019	698	Sac I	Bam HI	4	100
6	18016	18794	780	Bcl I	(Xho I)	5	60

Nucleotide positions are numbered according to GenBank entry AY188393. Restriction sites that were used for sub-cloning the assembled fragments into the expression vectors are bracketed.
^*Start and end positions include the corresponding restriction endonuclease recognition sites.
^**Following fragment assembly, clones were re-sequenced using specific primers; no mutations were found.
RE restriction endonuclease.

ISSN: 1756-0500