Cloning the Vector
DNA binding domain vector: pOBD2 (with uniques restriction sites and main features)
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Sequence features:
• ADH1 promoter: 1-298 (at least; region of homology when searched against SGD) • GAL4-DBD: 434-982 • ADH1 terminator: 1221-1687 (at least; region of homology to SGD genom. seq.) • beta lactamase (amp resistance): 4363-5220 • TRP1: 3379-4050 • CEN4: 1814-2667 Cloning site (position 877-1012): C TAT CTA TTC GAT GAT GAA GAT ACC CCA CCA AAC CCA AAA AAA GAG ATC GAA TTC CAG CTG ACC ACC ATG GCA ATT CCC GGG GAT CCG TCG ACC TGC AGA GAT CTA TGA ATC GTA GAT ACT GAA AAA CCC CGC AAG This sequence corresponds exactly to the two re-PCR primers (green + blue = forward primer, pink + black = reverse primer). The two restriction sites for linearizing pOBD2 (PvuII = CAGCTG and NcoI = CCATGG) are underlined. In the 4 bp-region of mixed color (blue and pink) the two primer sequences overlap). |
