pRS319a (ATCC® 87556)

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

Permits and Restrictions

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Designations pRS319a
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): 10.3059997558593800
Vector: pRS319a (phagemid)
Promoters: Promoter for in vitro transcription T7
Construction: pRS315
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
YC-type (centromeric) shuttle vector
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.

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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component of:ATCC 87562
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