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Oligos applied in this study. I: Synthesized DNA templates bearing either wild-type binding site (Zif_1) for zif268 or one of its variants (Zif_2 to Zif_6) used for generating DNA binding sites by PCR amplification, where KS-1 and SK-1 are two primers (low case). II: Oligos employed to construct five zif268 variants with QuickChange™ XL site-directed mutagenesis Kit (Stratagene) using pzif268 as a template. |
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| I |
Zif_1 tcgaggtcgacggtatcGCGTGGGCGCtccactagttctagagcggccgccac |
| Zif_2 tcgaggtcgacggtatcGCGTGGGCACtccactagttctagagcggccgccac |
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| Zif_3 tcgaggtcgacggtatcGCGTGGGCCCtccactagttctagagcggccgccac |
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| Zif_4 tcgaggtcgacggtatcGCGTGGGAGCtccactagttctagagcggccgccac |
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| Zif_5 tcgaggtcgacggtatcGCGTGGGAACtccactagttctagagcggccgccac |
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| Zif_6 tcgaggtcgacggtatcGCGTGGGACCtccactagttctagagcggccgccac |
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| KS-1 tcgaggtcgacggtatc |
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| SK*-1 gtggcggccgctctagaact (SK-1 was fluorescent labeled with either FAM, HEX, TAMRA, ROX, or CY5) |
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| II |
18Q_plus 5' CGCCGCTTTTCTcagTCGGATGAGCTTACCCGCC |
| 18Q_minus 5' GGCGGGTAAGCTCATCCGActgAGAAAAGCGGCG |
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| 18D_plus 5' CGCCGCTTTTCTgatTCGGATGAGCTTACCCGCC |
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| 18D_minus 5' GGCGGGTAAGCTCATCCGAatcAGAAAAGCGGCG |
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| 21N_plus 5' CGCCGCTTTTCTCGCTCGGATaacCTTACCCGCC |
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| 21N_minus 5' GGCGGGTAAGgttATCCGAGCGAGAAAAGCGGCG |
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| 18Q_21N_plus 5' CGCCGCTTTTCTcagTCGGATaacCTTACCCGCC |
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| 18Q_21N_minus 5' GGCGGGTAAGgttATCCGActgAGAAAAGCGGCG |
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| 18D_21N_plus 5' CGCCGCTTTTCTgatTCGGATaacCTTACCCGCC |
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| 18D_21N_minus 5' GGCGGGTAAGgttATCCGAatcAGAAAAGCGGCG |
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Liu and Stormo BMC Bioinformatics 2005 6:176 doi:10.1186/1471-2105-6-176 |
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