TABLE 2.

Oligonucleotide primers used for cloning of chromosomal fragments and detection of gene replacement

Primer purpose and primer	Comment	Sequencea
For cloning
B5	fxuB forward primer	5′-ACGAATTCACACGCCAGAGAACGC-3′
B6	fxuB reverse primer	5′-CAGGATCCTGATCTGGCAATTCCAG-3′
B7	fxbA upstream reverse primer	5′-CAGGATCCATTGGTAAGCCTTACC-3′
B8	fxbA upstream forward primer	5′-GATCTAGAGCTGGTTCAGGGCGATG-3′
U1	fxuC forward primer	5′-ACGAATTCGAGATCCTCGGCATCAAC-3′
U5	fxuC reverse primer	5′-CAGGATCCTGGCGCTTCCCGAAATC-3′
U6	fxuB forward primer	5′-GAGGATCCTGAAAGAACATATGACTC-3′
U7	fxuB reverse primer	5′-TATCTAGACAAGATCGGCCGACATA-3′
For detection of gene replacement
K1	Out of 5′ end of aph	5′-GAGACACAACGTGGCTTTC-3′
K2	Out of 3′ end of aph	5′-CACGAGGCAGACCTCAGC-3′
B1	Upstream of B8	5′-GGTACCGACACCTCGCA-3′
B9	Out of 3′ end of fxuA	5′-GGCGGGATCTACCTGGTAT-3′
U8	fxuC, upstream of U1 5′ end of fxb	5′-TCCAGGCCGTGGGACGTAA-3′
U9		5′-GTCGATGATCTGGCAATTCC-3′

^aRestriction sites are in bold face.