Size (kb): 10.8999996185302700
Vector: pHK9 (plasmid)
Construct size (kb): 10.89999961853027
Features: marker(s): ampR, LYS2
MCS: SalI (delta15)
restriction site: EcoRI,BamHI,XhoI,HpaI,NruI,BglII,PstI
restriction site: PvuII
coding sequence: karldelta15
Restriction digests of the clone give the following sizes (kb): BamHI--10.9; EcoRI--6.5, 2.4, 2.05; PstI--9.0, 1.8; PvuII--10.9.
Restriction digests of the clone give the following sizes (kb): BglII--7.8; EcoRI--7.8; HindIII--5.2, 2.5; EcoRI/HindIII--5.2, 2.5.
Supports introduction of kar1 delta 15 mutation into lys2 yeast strains by two step gene replacement. The karl delta 15 mutation supports transfer of YAC clones between yeast strains by Karl-mating (REF).
To integrate pHK9, linearize within the kar1 gene at the unique PvuII site. Cells that have lost the LYS2 marker through mitotic excision can be selected on alpha- aminoadipate.
Vallen EA, et al. Separate domains of KAR1 mediate distinct functions in mitosis and nuclear fusion. J. Cell Biol. 117: 1277-1287, 1992. PubMed: 1607389
Spencer F, et al. Yeast kar1 mutants provide an effective method for YAC transfer to new hosts. Genomics 22: 118-126, 1994. PubMed: 7959757
Hugerat Y, et al. A versatile method for efficient YAC transfer between any two strains. Genomics 22: 108-117, 1994. PubMed: 7959756
Spencer F, Simchen GTransfer of YAC clones to new hosts by karyogamy deficient mating.In: Spencer F, Simchen GCurrent Protocols in Human GeneticsNew York, N.Y.John Wiley and Sonsin press, 1995