pRS315 (ATCC® 77144)

Applications: YC-type (centromeric) shuttle vectorshuttle vectorvector containing primer sites useful for sequencingvector permitting RNA synthesis in vitrovector permitting production of single-stranded DNAvector permitting visual detection of recombinants  /  Depositors: P Hieter

Designations pRS315
Depositors P Hieter
Biosafety Level 1
Vector Information
Size (kb): 6.018
Vector: pRS315 (phagemid)
Promoters: Promoter for in vitro transcription T7
Construction: pRSS56 [pBluescript KS+, pBS(+)]
Marker(s):LEU2,ampR
Construct size (kb): 6.018
Features: insert detection: lacZ'
marker(s): LEU2
marker(s): ampR
promoter: lac
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7
replicon: ARSH4
replicon: f1
replicon: pMB1
MCS: SacI...KpnI
centromere: CEN6
Applications
YC-type (centromeric) shuttle vector
shuttle vector
vector containing primer sites useful for sequencing
vector permitting RNA synthesis in vitro
vector permitting production of single-stranded DNA
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--3.9, 2.1; BamHI--6.0; HindIII--6.0; KpnI--4.3, 1.7; KpnI/EcoRI--4.1, 1.5, 0.4.
One of a series of pBluescript-based centromere vectors (ATCC 77142-77145, 77157-77158) differing in the yeast selectable marker gene.
YC-type centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ') peptide.
pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and f1 origin of pBS(+).
A fragment (2.235 kb) containing the LEU2 gene was inserted into the NdeI site and a cassette containing CEN6 and the ARS associated with histone 4 (ARSH4) was inserted into the AatII site of pRSS56. All ends were blunted.
The order of the major features in this plasmid is: LEU2 - f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19-27, 1989. PubMed: 2659436

Related Products
component of:ATCC 77190
Shipped frozen
Shipping Information Freeze dried E. coli containing the phagemid.
Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 77144