M3499 [URA3 --> ADE2 converter] (ATCC® 87558)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector URA3 --> ADE2  /  Depositors: DJ Stillman

Designations M3499 [URA3 --> ADE2 converter]
Permits and Restrictions

View Permits

Depositors DJ Stillman
Biosafety Level 1
Vector Information
Size (kb): 7.4260001182556150
Vector: M3499 (plasmid)
Construction: pUC18
Marker(s):ampR ADE2
Construct size (kb): 7.426000118255615
Features: gene disruption cassette: ura3::ADE2
marker(s): ampR
replicon: pMB1
restriction site: HindIII
restriction site: HindIII...EcoRI
restriction site: PstI
YI-type (integrating) shuttle vector
host modification
marker swap vector URA3 --> ADE2
Restriction digests of the clone give the following sizes (kb): HindIII--2.5, 1.3, 1.1, 1.1, 1.0; BamHI--6.6, 0.5; PvuI--6.2, 0.8.
To convert the host phenotype from URA3 to ADE2, transform with the PstI digested vector (4.45 kb) and select for Ade+ transformants.
A marker swap (URA3->ADE2) vector for altering a Saccharomyces cerevisiae host by a one-step gene disruption.
When swapping markers on an episomal plasmid, appropriate phenotype may result from loss of the plasmid unless a second selectable or scorable marker is used to ensure plasmid maintenance.
Vector was constructed by replacing an internal StuI-EcoRV fragment of URA3 with a 3.7 kb BamHI fragment containing the ADE2 coding sequence. ADE2 and URA3 are in the same orientation.
Growth Conditions
Temperature: 37.0°C

Kasten MM, et al. A large protein complex containing the yeast Sin3p and Rpd3p transcriptional regulators.. Mol. Cell. Biol. 17(8): 4852-4858, 1997. PubMed: 9234741

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