pRS319a (ATCC® 87556)

Applications: YC-type (centromeric) shuttle vector  /  Depositors: DJ Stillman

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Designations pRS319a
Depositors DJ Stillman
Biosafety Level 1
Vector Information
Size (kb): 10.3059997558593800
Vector: pRS319a (phagemid)
Promoters: Promoter for in vitro transcription T7
Construction: pRS315
Marker(s):LEU2,ampR,CAN1
Construct size (kb): 10.30599975585938
Features: marker(s): CAN1
marker(s): LEU2
marker(s): ampR
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7
replicon: ARS4
replicon: f1
replicon: pMB1
MCS: SacI...KpnI
centromere: CEN6
Applications
YC-type (centromeric) shuttle vector
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--4.1, 2.8, 2.6, 1.1; HindIII--6.6, 2.8, 1.4; PstI--9.5, 1.0.
pRS319a differs from pRS319 in that pRS319 has a truncated CAN1 gene and pRS319a has a full length CAN1 gene.
A plasmid shuffling vector. YC-type centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains CAN1 marker (full length CAN1 gene) for negative selection.
pRS319a was constructed by inserting a 4.161 kb PvuII-Sma CAN1 fragment from M3093 (CAN1 in pBluescript II) into the HpaI site (position 2175) of pRS315 (ATCC 77144).
The order of the major features in this plasmid is: LEU2 - CAN1 - f1 ori - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4.
References

Sikorski RS, Boeke JD. In vitro mutagenesis and plasmid shuffling: from cloned gene to mutant gene. Methods Enzymol. 194: 302-318, 1991. PubMed: 2005795

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