M2371 [HIS3 --> ADE2 converter] (ATCC® 87557)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector HIS3 --> ADE2  /  Depositors: DJ Stillman

Designations M2371 [HIS3 --> ADE2 converter]
Permits and Restrictions

View Permits

Depositors DJ Stillman
Biosafety Level 1
Vector Information
Size (kb): 7.7340002059936520
Vector: M2371 (plasmid)
Construction: pUC18
Marker(s):ampR ADE2
Construct size (kb): 7.734000205993652
Features: gene disruption cassette: his3::ADE2
marker(s): ampR
replicon: pMB1
restriction site: BamHI...EcoRI
restriction site: HindIII...BamHI
YI-type (integrating) shuttle vector
host modification
marker swap vector HIS3 --> ADE2
Restriction digests of the clone give the following sizes (kb): BamHI--3.7, 2.7, 0.7, 0.6; EcoRI--6.8, 0.9; PvuI--6.8, 0.9; HindIII--3.9, 1.6, 1.2, 1.0.
To convert the host phenotype from HIS3 to ADE2, transform with the PstI+SacI digested vector (4.99 kb) and select for Ade+ transformants.
A marker swap (HIS3->ADE2) vector for altering a Saccharomyce s 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 MscI-NsiI fragment of HIS3 with a 3.7 kb NotI (blunt)-PstI fragment containing the ADE2 coding sequence. ADE2 and HIS3 are in the opposite orientation.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C

Macatee T, et al. Global alterations in chromatin accessibility associated with loss of SIN4 function.. Nucleic Acids Res. 25(6): 1240-1247, 1997. PubMed: 9092635

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