ATCC 100 Years Logo Anniversary ATCC 100 Years Logo Anniversary Cart 0
  • Careers
  • Support

pDA71

77474

Organism
Escherichia coli (Migula) Castellani and Chalmers
Classification
Enterobacteriaceae, Escherichia
Clone type
Vector
Applications
Molecular biology
Product format
Frozen
Mission Collection Item
This is a Mission Collection Item.

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
contains sequence chloramphenicol resistance (non-enzymatic)
contains sequence restriction endonuclease EcoRI
in another host, produces protein chloramphenicol resistance (non-enzymatic)
produces protein restriction endonuclease EcoRI
shuttle vector
vector for constructing genomic libraries
vector permitting positive selection for inserts

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--5.4, 3.3; BglII--8.8; HindIII--8.8.
Cloning sites are positioned in the EcoRI endonuclease gene, regulated by a lambda promoter. In E. coli hosts providing the temperature-sensitive lambda repressor cI857, growth at 37C will occur only if the endonuclease gene is interrupted.
When using the positive selection feature, the E. coli recipient should not be a lambda lysogen because the feature is turned off by a lambda repressor.
The non-enzymatic chloramphenicol resistance gene, derived from Rhodococcus rhodochrous SC2318 (ATCC 12674), does not confer resistance in E. coli.
The plasmid confers immunity to nocardiophage Q4.
Utilization of the positive selection feature of this vector requires growth (37C, 50 ug/ml ampicillin) in hosts not expressing cI857, before transformation into nocardioforms.
Subsequent large-scale growth of this stock (100 ug/ml ampicillin, 30 ug/ml rifampicin), allows preparation of enough plasmid DNA to transform the nocardioform recipient.
Libraries should be allowed to grow in nocardioforms at 26 degrees for 12 hours before introducing selection. Chloramphenicol should be applied underneath the agar and be allowed to diffuse to a concentration of 40 ug/ml.
Shuttle vector permitting positive selection in E. coli.
E. coli MM294-1(lambda) is a rifampicin resistant derivative of E. coli MM294.
The order of the major features in the plasmid is: AccIII - phage Q4 replicon - StuI - Cm-R gene - BbrPI - BbrPI - AccIII - EcoRI gene - ampR.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
8.800000190734863
Vector name
pDA71 (plasmid)
Type of vector
plasmid
Construction
Cm-R#17, pEcoR251
Insert detection
hsdR
Markers
cmlR; ampR
Promoters
lambda
Replicon
pMB1; phage Q4

Insert information

Type of DNA
genomic
Target gene
restriction endonuclease EcoRI, chloramphenicol resistance (non-enzymatic)
Gene product
chloramphenicol resistance (non-enzymatic) [Cm-R]

Handling information

History

Depositors
ER Dabbs

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

Quan S, Dabbs ER. Nocardioform arsenic resistance plasmid characterization and improved Rhodococcus cloning vectors. Plasmid 29: 74-79, 1993. PubMed: 8441772

Eric R Dabbs, personal communication

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

Message Us

Hours of Operation

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