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To replace 704-1055 with URA3 marker from pRS406
- SPT6 from 704-744: 5'- CTGGTCTATCGTCGGATAAGATTGACGAGATGTATGACAT
- fwd URA3: 5' AGATTGTACTGAGAGTGCAC
- SPT6 from 1015-1055: 5' ATGTCATCGGAGGATCAAGAATTAGAAAGAAACTGGATAG
- rev complement SPT6 (from 1055-1015): 5'- 5'- CTA TCC AGT TTC TTT CTA ATT CTT GAT CCT CCG ATG ACA T
- rev URA3: 5' CTGTGCGGTATTTCACACCG
5' CTGTGCGGTATTTCACACCGCTA TCC AGT TTC TTT CTA ATT CTT GAT CCT CCG ATG ACA T
- stepwise move centromere from one end of chromosome to other
- progressively larger deletions from telomere to first essential gene on each chromosome
- clustering highly expressed genes on one arm, poorly expressed genes on other
- clustering related genes into "operon"
- multiple copies of yeast mating locus
- replace S. cerevisiae genes for single complex (e.g. SAGA) with homologs from other yeast starting with pombe
- in vivo reporter for transciption elongation: functional RNA?
- randomize sigma 4.2 region of TFG2 and screen library for rifS and FLO:HIS mutants
- check Drew's T7.1 for revertants that increase plaque size back to normal.
- biobrick assembly for yeast parts (look at Pam's yeast parts first!)...ideally would use homologous recomb vs restriction sites