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Primer sequences used to amplify intergenic fragments for subsequent cloning |
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| Construct* |
Primer sequence |
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| (Forward) |
(Reverse) |
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| pGL3-Basic constructs |
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| 1. SERPINI1 (1-851) |
5'-ATAGATCTACTCCGGCGACGCCGGA-3' |
5'-ATAAGCTTGTCCAGACTGCGCCTCT-3' |
| 2. SERPINI1 (1-175) |
5'-ATAGATCTACTCCGGCGACGCCGGA-3' |
5'-ATAAGCTTTCGCCTCAGGATCCCCC-3' |
| 3. SERPINI1 (176-851) |
5'-CGAGATCTTGGGTGGTGACACTTCT-3' |
5'-ATAAGCTTGTCCAGACTGCGCCTCT-3' |
| 4. SERPINI1 (474-851) |
5'-AGAGATCTGTGTGGGGACTTGCACT-3' |
5'-ATAAGCTTGTCCAGACTGCGCCTCT-3' |
| 5. SERPINI1 (711-851) |
5'-ATAGATCTCAGGGCAGCTGCTTTCC-3' |
5'-ATAAGCTTGTCCAGACTGCGCCTCT-3' |
| 6. PDCD10 (851-1) |
5'-ATAGATCTGTCCAGACTGCGCCTCT-3' |
5'-ATAAGCTTACTCCGGCGACGCCGGA-3' |
| 7. PDCD10 (710-1) |
5'-TCAGATCTGTACCTTTCCGCTCCCT-3' |
5'-ATAAGCTTACTCCGGCGACGCCGGA-3' |
| 8. PDCD10 (473-1) |
5'-ATAGATCTCAAGCCCTCCCTTCACC-3' |
5'-ATAAGCTTACTCCGGCGACGCCGGA-3' |
| 9. PDCD10 (175-1) |
5'-ATAGATCTTCGCCTCAGGATCCCCC-3 |
5'-ATAAGCTTACTCCGGCGACGCCGGA-3' |
| 10. PDCD10 (107-1) |
5'-ATAGATCTGCCGGCGAGCGGGACTA-3' |
5'-ATAAGCTTACTCCGGCGACGCCGGA-3' |
| 11. PDCD10 (175-108) |
5'-ATAGATCTTCGCCTCAGGATCCCCC-3' |
5'-CCAAGCTTGTTGTCCGCAATTCCTG-3' |
| 12. PDCD10 (851-176) |
5'-ATAGATCTGTCCAGACTGCGCCTCT-3' |
5'-CGAAGCTTTGGGTGGTGACACTTCT-3' |
| Heterologous promoter constructs |
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| 1. SERPINI1 (176-851)/S |
5'-CCTGCTAGCTGGGTGGTGACACTTCTAG-3' |
5'-AAAGATCTGTCCAGACTGCGCCTCTG-3' |
| 2. PDCD10 (851-176)/S |
5'-TAAGCTAGCGTCCAGACTGCGCCTCTG-3' |
5'-CCTAGATCTTGGGTGGTGACACTTCTAG-3' |
| 3. PDCD10 (473-176)/S |
5'-TATGCTAGCCAAGCCCTCCCTTCACCTGG-3' |
5'-CCTAGATCTTGGGTGGTGACACTTCTAG-3' |
| 4. PDCD10 (710-474)/S |
5'-GCTGCTAGCGTACCTTTCCGCTCCCTG-3' |
5'-GAAGATCTGTGTGGGGACTTGCACTG-3' |
| 5. PDCD10 (851-711)/S |
5'-TAAGCTAGCGTCCAGACTGCGCCTCTG-3' |
5'-TTAGATCTCAGGGCAGCTGCTTTCCG-3' |
| 6. SERPINI1 (176-473)/C |
5'-CCTGCTAGCTGGGTGGTGACACTTCTAG-3' |
5'-CCAGATCTCAAGCCCTCCCTTCACCTGG-3' |
| 7. SERPINI1 (474-710)/C |
5'-GGAGCTAGCGTGTGGGGACTTGCA-3' |
5'-CTAGATCTGTACCTTTCCGCTCCCTG-3' |
| 8. SERPINI1 (711-851)/C |
5'-CTTGCTAGCCAGGGCAGCTGCTTTCCG-3' |
5'-AAAGATCTGTCCAGACTGCGCCTCTG-3' |
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*Construct names with an "S" character at the end indicates that the PCR-amplified intergenic product was cloned into the upstream of SV40 minimal promoter. Construct names with a "C" character denotes that the amplified intergenic segment was cloned into the upstream of CMV promoter. | ||
Chen et al. BMC Molecular Biology 2007 8:2 doi:10.1186/1471-2199-8-2 |
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