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

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

Permits and Restrictions

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Designations M2371 [HIS3 --> ADE2 converter]
Depositors DJ Stillman
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

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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Shipped freeze-dried