pTL7 [TRP1 --> LEU2 converter] (ATCC® 87549)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector TRP1 --> LEU2  /  Depositors: FR Cross

Designations pTL7 [TRP1 --> LEU2 converter]
Permits and Restrictions

View Permits

Depositors FR Cross
Biosafety Level 1
Vector Information
Size (kb): 7.8000001907348630
Vector: pTL7 (phagemid)
Construction: pBC KS+
Construct size (kb): 7.800000190734863
Features: gene disruption cassette: tr:LEU2/kanR
marker(s): cmlR
replicon: f1
replicon: pMB1
restriction site: SmaI
restriction site: XhoI
YI-type (integrating) shuttle vector
host modification
marker swap vector TRP1 --> LEU2
Restriction digests of the clone give the following sizes (kb): EcoRI--6.2, 1.7; HindIII--7.8; SmaI/XhoI--4.4, 3.3.
To convert the host phenotype from TRP1 to LEU2, transform with the XhoI+SmaI digested vector and select for Leu+ transformants.
Some combinations of marker swap plasmids and target locus may result in relatively high reversion rates. In most but not all cases the frequencies of successful convertants are greater than 30%.
A marker swap vector designed to change the S. cerevisiae host phenotype by one-step gene disruption of the TRP1 gene with the LEU2 and kanR markers.
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 EcoRV fragment of TRP1 with a SmaI fragment containing the LEU2 and kanR coding sequences. LEU2 and TRP1 are in the same orientation.
Media ATCC Medium 1065 (see below) plus kanamycin (50.0 mcg/ml) plus chloramphenicol (25.0 mcg/ml) ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0 L
Growth Conditions
Temperature: 37.0°C

Cross FR. 'Marker swap' plasmids: convenient tools for budding yeast molecular genetics. Yeast 13: 647-653, 1997. PubMed: 9200814

Related Products
component of:ATCC 87561
Shipped freeze-dried