pSVA3

77273 ^™

This clone encodes the L22F mutant of DHFR that can serve as a dominant selection marker because it retains catalytic activity and also confers methotrexate (MTX) resistance at low MTX concentrations (125 nM). Transfected cells amplify DHFR sequences. Constructed by cloning a HindIII/SmaI fragment containing the gene into pSV2-neo from which neoR had been removed. This plasmid was then cut with SmaI and BglII to remove 18 G nucleotides at the 3? end. The BglII site was filled in, and ligated to the SmaI site, thereby destroying the original BglII and SmaI sites. Removing the HindIII/SmaI fragment from the pSV2-neo removes all but the 3? 171 bp of neoR.

Organism: Cricetulus griseus, hamster, Chinese
Clone type: Clone
Applications: Molecular biology
Product format: Freeze-dried
Storage conditions: 2°C to 8°C

See Additional Product Information

Price: $639.00 EA

Discounts may be available for our fellow nonprofit organizations. Login to see your price.

Generally ships within 1-3 business days

Select specific lot(s)

Please enter the quantity for the selected lot number and add to cart.

Please fill at least one quantity field.

Quantity should not be greater than inventory.

Quantity	Lot Number	Inventory	Donor Age	Donor Sex	Donor Ethnicity	Donor BMI	CoA

Documentation

Product sheet

BSL 1

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.

General

Specific applications: in another host, produces protein dihydrofolate reductase

Characteristics

Comments: Restriction digests of the clone give the following sizes (kb): BamHI--3.5, 1.7; EcoRI--2.7, 2.3; PstI--1.9, 1.4, 0.95, 0.9; PvuII--4.0, 1.1; HindIII/SmaI--5.2.
Transfected cells amplify DHFR sequences.
Encodes the L22F mutant of DHFR that can serve as a dominant selection marker because it retains catalytic activity and also confers methotrexate (MTX) resistance at low MTX concentrations (125 nM).
Constructed by cloning a HindIII/SmaI fragment containing the gene into pSV2-neo from which neoR had been removed. This plasmid was then cut with SmaI and BglII to remove 18 G nucleotides at the 3' end.
The BglII site was filled in, and ligated to the SmaI site, thereby destroying the original BglII and SmaI sites.
Removing the HindIII/SmaI fragment from pSV2-neo removes all but the 3' 171 bp of neoR.
Mycoplasma contamination: Not detected

Vector information

Construct size (kb): 5.099999904632568
Intact vector size: 5.729
Vector name: pSV2-neo
Type of vector: plasmid
Construction: SV40, Tn5, pBR322
Host range: vertebrate cells
Vector end: HindIII; SmaI
Vector information: Cross references: DNA Seq. Acc.: U02434
Cloning sites: EcoRI; BamHI; PvuII; PstI; HindIII
Markers: ampR
Promoters: SV40 early
Replicon: SV40; pMB1
Restriction sites: HindIII

Insert information

Insert size (kb): 0.70999999999999996
Type of DNA: cDNA
Insert source: lung cell line
Insert tissue: lung cell line
Insert information: DESCRIPTION OF INSERT COMPONENT:
Insert 5' end: HindIII
Insert 3' end: BglII
Cross references:
Genome: hamster, Chinese
Target gene: dihydrofolate reductase
Gene name: dihydrofolate reductase
Gene product: dihydrofolate reductase [DHFR]
Gene symbol: DHFR
Contains complete coding sequence: Unknown

Handling information

Host: Escherichia coli HB101 (ATCC 33694)
Medium: ATCC Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Temperature: 37°C
Handling procedure: Transfer a loopful to a test tube containing 5 mL LB+50mg/mL of ampicillin broth. A loopful of culture can also be streaked on an LB + amp agar plate. Incubate cultures at 37°C. Isolate DNA using standard plasmid preparation procedures.
Handling notes: Restriction digests of the clone gave the following sizes (in kb): HindIII/SmaI -- 5.2; BamHI -- 3.5, 1.7 ; EcoRI -- 2.7, 2.3 ; PstI -- 1.9, 1.4, 0.95, 0.9 ; PvuII -- 4.0, 1.1. ---ATCC staff
Restriction digests of the clone give the following sizes (kb): BamHI--3.5, 1.7;
EcoRI--2.7, 2.3; PstI--1.9, 1.4, 0.95, 0.9; PvuII--4.0, 1.1; HindIII/SmaI--5.2.
- ATCC staff

Encodes the L22F mutant of DHFR that can serve as a dominant selection marker
because it retains catalytic activity and also confers methotrexate (MTX)
resistance at low MTX concentrations (125 nM).
- Gene 112: 179-188, 1992

Transfected cells amplify DHFR sequences.
- Gene 112: 179-188, 1992

Constructed by cloning a HindIII/SmaI fragment containing the gene into pSV2-neo
from which neoR had been removed. This plasmid was then cut with SmaI and BglII
to remove 18 G nucleotides at the 3' end.
- Gene 112: 179-188, 1992

The BglII site was filled in, and ligated to the SmaI site, thereby destroying
the original BglII and SmaI sites.
- Gene 112: 179-188, 1992

Removing the HindIII/SmaI fragment from pSV2-neo removes all but the 3' 171 bp
of neoR.
- Gene 112: 179-188, 1992

History

Depositors: PW Melera

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

References

Curated Citations

Hussain A, et al. Construction of a dominant selectable marker using a novel dihydrofolate reductase. Gene 112: 179-188, 1992. PubMed: 1555767

For product-related inquiries and issues, contact Product Experience:

Message Us

Send us a message

Hours of Operation

Monday - Friday

9:00am - 5:00pm
US Eastern Time