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pdeltaADE2

99604

Clone type
Vector
Applications
Molecular biology
Product format
Frozen
Mission Collection Item
This is a Mission Collection Item

Documentation

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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
contains sequence ATP phosphoribosyltransferase
marker deletion vector phosphoribosylaminoimidazole succinocarboxamide synthetase SAICAR synthetase, phosphoribosylaminoimidazole carboxylase
produces protein uridine monophosphate synthetase UMP synthase, orotate phosphoribosyltransferase, orotidine 5'-phosphate decarboxylase, orotate phosphoribosyltransferase 1

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): BamHI--5.2, 3.2; EcoRI--5.0, 3.4; HindIII--7.0, 1.3.
The two step selection process requires a ura3 transformation host (this host can be created using pJL164 (ATCC 87471)). After transformation with the BamHI digested plasmid, URA3 integrants are selected on ura- plates.
The designer deletion strain is then recovered by selection on 5-FOA plates (loss of URA3 and ADE2 markers by a homologous recombination event between the two hisG repeats).
The deleted host retains the coding sequence for six C-terminal amino acids of ADE2.
E. coli containing this plasmid should be grown on media lacking pyrimidines to select for URA3-containing cells.
This deleter vector is used to create designer yeast strains with a non-revertable ade2 auxotrophic marker deletion.
The 5.2 kb BamHI insert contains two direct repeats of the Salmonella hisG gene flanking URA3 and about 700 bp of homology to sequences upstream and downstream of the ADE2 gene flanking the hisG-URA3-hisG sequence.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
8.4
Intact vector size
8.400
Vector name
pdeltaADE2 (plasmid)
Type of vector
plasmid
Construction
pBluescript, URA3, hisG, ADE2 sequences
Host range
Saccharomyces cerevisiae
Escherichia coli
Vector information
other: ADE2 flanking sequence
other: ADE2 flanking sequence
other: ADE2 flanking sequence
Coding sequence
hisG, ->; hisG, ->; hisG
Markers
ampR; URA3
Replicon
pMB1
Restriction sites
BamHI

Insert information

Target gene
ATP phosphoribosyltransferase; uridine monophosphate synthetase

Handling information

Host
Escherichia coli MC1066
Medium
Temperature
37°C
Handling notes
Restriction digests of the clone give the following sizes (kb): BamHI--5.2, 3.2;
EcoRI--5.0, 3.4; HindIII--7.0, 1.3.
- ATCC staff

E. coli containing plasmid should be grown on medium lacking pyrimidines to
select for URA3-containing cells.
- personal communication

The 5.2 kb BamHI insert contains two direct repeats of the Salmonella hisG gene
flanking URA3 and about 700 bp of homology to sequences upstream and downstream
of the ADE2 gene flanking the hisG-URA3-hisG sequence.
- Cell 66: 1279-1287, 1991

This deleter vector is used to create designer yeast strains with a
non-revertable ade2 auxotrophic marker deletion.
- Cell 66: 1279-1287, 1991

The two step selection process requires a ura3 transformation host (this host
can be created using pJL164 (ATCC 87471)). After transformation with the BamHI
digested plasmid, URA3 integrants are selected on ura- plates.
- Cell 66: 1279-1287, 1991

The designer deletion strain is then recovered by selection on 5-FOA plates
(loss of URA3 and ADE2 markers by a homologous recombination event between the
two hisG repeats).
- Cell 66: 1279-1287, 1991

The deleted host retains the coding sequence for six C-terminal amino acids of
ADE2.
- Cell 66: 1279-1287, 1991

History

Depositors
JD Boeke
Special collection
NCRR Contract

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.
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Disclaimers

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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.

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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

References

Curated Citations

Aparicio OM, et al. Modifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae. Cell 66: 1279-1287, 1991. PubMed: 1913809

Alani E, et al. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics 116: 541-545, 1987. PubMed: 3305158

Jef D Boeke, personal communication

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