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Distribution host: Escherichia coli HB101 (ATCC 33694)

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

shuttle vector

vector permitting visual detection of recombinants

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.

Temperature: 37.0°C