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Table 1 Primer sequences used to amplify intergenic fragments for subsequent cloning

From: Two non-homologous brain diseases-related genes, SERPINI1 and PDCD10, are tightly linked by an asymmetric bidirectional promoter in an evolutionarily conserved manner

Construct*

Primer sequence

 

(Forward)

(Reverse)

pGL3-Basic constructs

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

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'

  1. *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.
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BMC Molecular Biology

ISSN: 1471-2199

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