pRC2312 (ATCC® 77411)

Applications: shuttle vectorvector permitting visual detection of recombinants  /  Depositors: RD Cannon

Permits and Restrictions

View Permits

Designations pRC2312
Depositors RD Cannon
Biosafety Level 1
Host
Distribution host: Escherichia coli HB101 (ATCC 33694)
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
Applications
shuttle vector
vector permitting visual detection of recombinants
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.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
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

Shipped frozen
Shipping Information Distributed: freeze-dried
Product Sheet
Product Sheet
Product Sheet