Restriction digests of the clone give the following sizes (kb): BamHI--5.3; XbaI--5.3; PstI--5.3; HindIII--5.3.
This is a Escherichia coli/yeast episomal shuttle vector with multiple restriction sites for cloning.
Constructed by filling in and blunt-end ligating pUC18 linearized at NdeI, a 1.4 kb RsaI fragment of the 2 micron circle, and a 1.1 kb HindIII/SmaI fragment containing URA3. A XbaI site in 2u and a PstI site in URA3 were removed.
The yeast genomic sequence is in nt 286-1383 (where the A of the ATG of lacZ' is number 1), the 2 micron sequence goes from nt 1384-2768, and pUC18 includes nt 2769-285. |
