User:Nkuldell/other rewrites

spt6:URA3

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

spt6:URA3_704_1055_fwd

5' CTGGTCTATCGTCGGATAAGATTGACGAGATGTATGACATAGATTGTACTGAGAGTGCAC

spt6:URA3_704_1055_rev

5' CTGTGCGGTATTTCACACCGCTA TCC AGT TTC TTT CTA ATT CTT GAT CCT CCG ATG ACA T

ideas

• 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