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pRS401 plasmid in E. coli

87473

Clone type
Vector
Applications
Molecular biology
Product format
Frozen
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Detailed product information

General

Specific applications
YI-type (integrating) shuttle vector
vector permitting visual detection of recombinants O-acetylhomoserine sulfhydrylase

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): BamHI--4.9; EcoRI--3.2, 1.7; XbaI--2.9, 2.0.
met15 phenotype produces brown colonies when grown on Pb containing media.
This requires two approx. 60nt PCR primers; the 20nts of sequence at the 3' ends of each primer is specific for amplifying the MET15 gene from pRS401, and the 40nts of sequence at the 5' ends matches the genomic sequences flanking the gene of interest.
These same primers can be used to amplify an ADE2 marker gene disruption product from pRS402 (ATCC 87477).
MET15, MET17 and MET25 are synonymous.
pRS401 can be used to generate a gene specific MET15 marker gene disruption cassette for tranformation in gene knockout experiments.
The 20nt PCR primer sequences for generating the MET15 marker from pRS401 are: 5'-CTGTGCGGTATTTCACACCG-3' (left primer) and 5'-AGATTGTACTGAGAGTGCAC-3' (right primer).
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
4.893
Intact vector size
4.893
Vector name
pRS401 (phagemid)
Type of vector
phagemid
Construction
pJK142
Host range
Saccharomyces cerevisiae
Escherichia coli
Cloning sites
SacI; BstXI; SacII; EagI; NotI; SpeI; BamHI; SmaI; PstI; ClaI; SalI; XhoI; ApaI; KpnI
Coding sequence
lacZ', <-, 2317-2676; lacZ'
Markers
MET15; ampR
MCS
KpnI...SacI, ->, 2513-2619
Polylinker sites
SacI; BstXI; SacII; EagI; NotI; XbaI; SpeI; BamHI; SmaI; PstI; EcoRI; EcoRV; HindIII; ClaI; SalI; XhoI; ApaI; KpnI
Promoters
In vitro transcription T7; lac
Replicon
f1, ←, 1860-2316; pMB1, 3075-3075

Insert information

Target gene
O-acetylhomoserine sulfhydrylase

Handling information

Host
Escherichia coli HB101 (ATCC 33694)
Medium
Temperature
37°C
Handling notes
Restriction digests of the clone give the following sizes (kb): BamHI--4.9;
EcoRI--3.2, 1.7; XbaI--2.9, 2.0.
- ATCC staff

MET15, MET17 and MET25 are synonymous.
- SGD

met15 phenotype produces brown colonies when grown on Pb containing media.
- Yeast 14: 115-132, 1998

pRS401 can be used to generate a gene specific MET15 marker gene disruption
cassette for tranformation in gene knockout experiments.
- Yeast 14: 115-132, 1998

This requires two approx. 60nt PCR primers; the 20nts of sequence at the 3' ends
of each primer is specific for amplifying the MET15 gene from pRS401, and the
40nts of sequence at the 5' ends matches the genomic sequences flanking the gene
of interest.
- Yeast 14: 115-132, 1998

The 20nt PCR primer sequences for generating the MET15 marker from pRS401 are:
5'-CTGTGCGGTATTTCACACCG-3' (left primer) and 5'-AGATTGTACTGAGAGTGCAC-3' (right
primer).
- Yeast 14: 115-132, 1998

These same primers can be used to amplify an ADE2 marker gene disruption product
from pRS402 (ATCC 87477).
- Yeast 14: 115-132, 1998

History

Depositors
JD Boeke
Special collection
NCRR Contract
Cross references
GenBank U93714

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Cost GJ, Boeke JD. A useful colony colour phenotype associated with the yeast selectable/counter-selectable marker MET15. Yeast 12: 939-941, 1996. PubMed: 8873447

Brachmann CB, et al. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14: 115-132, 1998. PubMed: 9483801

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