|
Oligonucleotide
|
Sequence (5’- 3’)a
|
Restriction site
|
Position of annealingb
|
|---|
|
cotS3
|
ggatccATCGACCATGTGGCGCTG
|
Bam HI
|
+64/+82
|
|
cotS4
|
aagcttAACCATCACTTTATTCAG
|
Hind III
|
+771/+758
|
|
cotZ1
|
gcatgcGCTGTTGAAGAAGACTGC
|
Sph I
|
+64/81
|
|
cotZ2
|
gtcgacAACTTCAATACAGTAGTTCG
|
Sal I
|
+378/+359
|
|
cgeA1
|
gcatgcCAACTGCAACAGAAGGAG
|
Sph I
|
+64/+81
|
|
cgeA2
|
gtcgacGTGAACCTGATCGAAAGC
|
Sal I
|
+297/+279
|
- a Capital and lowercase letters indicate nucleotides complementary to corresponding gene DNA and unpaired flanking sequences carrying a restriction site, respectively. b Referred to cotS, cotZ or cgeA sequences, taking the first nucleotide of the initiation codon as +1.
