| Vector Information |
Size (kb): 8.5800008773803710 DESCRIPTION OF VECTOR:
Intact vector size: 8.580
Type of vector: plasmid
Cloning sites: SmaI BamHI PstI HindIII
Polylinker sites: SmaI BamHI PstI HindIII
Construction: pUC9, URA3, LEU2, CaARS
Host range: Candida albicans; Saccharomyces cerevisiaeCandida robusta;
Escherichia coli
Features (with orientation and position when available):
insert detection: lac
marker(s): ampR, URA3, LEU2
promoter: lac
replicon: pMB1, CaARS Vector: pRC2312 (plasmid) Promoters: Promoter lac Construction: pUC9, URA3, LEU2, CaARS Marker(s):LEU2,URA3,ampR Construct size (kb): 8.580000877380371 Features: insert detection: lac
marker(s): ampR, URA3, LEU2
promoter: lac
replicon: pMB1, CaARS |
| Comments |
Restriction digests of the clone give the following sizes (kb): EcoRI--7.2, 1.2, 0.2; HindIII--8.6; BglI--7.0, 1.6; PvuI--6.8, 1.8; PvuII--8.3, 0.3. Transformation of S. cerevisiae AH22 with pRC2312 resulted in replicative transformants at high frequency (1.41(E)5 per ug DNA). Transformants of C. albicans SGY-243 with pRC2312 gave replicative transformants at high frequency (5.42(E)3 per ug DNA) and integrative transformants at lower frequency (32 per ug DNA). Shuttle vector maintained at approximately 15 copies per haploid genome in S. cerevisiae and 2-3 copies per genome in C. albicans. Recombinants can be detected using blue/white color selection. Candida albicans genes LEU2 and URA3 were isolated from plasmids pMK155 and pCARS1 respectively. The C. albicans autonomously replicating sequence (CaARS) allows replication in C. albicans and S. cerevisiae. The order of the major features in the plasmid is: ampR - pMB1 ori - lacZ/HindIII/MCS/SmaI - URA3 - CaARS - LEU2. |
| References |
Cannon RD, et al. Cloning and expression of Candida albicans ADE2 and proteinase genes on a replicative plasmid in C. albicans and in Saccharomyces cerevisiae. Mol. Gen. Genet. 235: 453-457, 1992. PubMed: 1465116
Cannon RD, et al. Isolation and nucleotide sequence of an autonomously replicating sequence (ARS) element functional in Candida albicans and Saccharomyces cerevisiae. Mol. Gen. Genet. 221: 210-218, 1990. PubMed: 2196431
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