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

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

Sub-fragment number	Start*	End*	Length	5′-flanking RE	3′-flanking RE	Clones sequenced**	Clones with the correct sequence, %
1	8532	9117	586	(Xho I )	Avr II	3	100
2	9114	9800	687	Xba I	Bam HI	3	67
3	9795	10573	779	Bam HI	Avr II	3	67
4	10570	11208	639	Spe I	Xho I	3	100
5	11205	12491	1287	Sal I	(Nco I)	4	100

Nucleotide positions are numbered according to the 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