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pGFLEX plasmid in E. coli

87629

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
Encodes removable tag for protein isolation
Expression vector
Shuttle vector
Vector permitting construction of fusion proteins
Vector permitting production of single-stranded DNA

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): PstI--6.2; XhoI--3.7, 2.0,0.3; SalI--2.4, 2.1, 1.6.

 

The pGFLEX expression system allows production of target proteins fused to either the N or C terminus of the GST pi protein and provides rapid purification of target proteins as either GST fusions or native proteins after cleavage with thrombin.

High level of expression in E. coli, cos and other mammalian cultured cell lines.
Recommendation for verification: StuI--4.84, 1.30; SalI--2.4, 2.18, 1.56; PstI--6.14; NotI--6.14.
Mycoplasma contamination
Not detected

Vector information

Intact vector size
6.143
Type of vector
phagemid
Construction
pUC19, pGPi-2, pIC-20H
Host range
mammalian cells
Vector information
Leader Peptide: beta-gal leader sequence, 848-886
Other: thrombin cleavage site, 1517-1524
Stop Codons: TGA, 1570-1572
Other unique sites: BglII BssHI EcoRV PstI PvuI ScaI SpeI
Cloning sites
ClaI; NotI; ApI
Coding sequence
human GST pi, ->, 887-1516; Neo, ->, 2836-3630
Markers
G418R; ampR
MCS
BamHI...ApaI, 1525-1566
Polylinker sites
BamHI; EcoRI; SmaI; SalI; XhoI; NotI; ApaI
Replicon
f1 ori, 1895-2413; SV40 ori, 2669-2754; pMB1, 4373-4373
Terminator
SV40 polyadenylation, 3804-3939

Handling information

Host
Escherichia coli HB101 (ATCC 33694)
Medium
Temperature
37°C
Handling notes
Restriction digests of the clone give the following sizes (kb): PstI--6.2; XhoI--4.1, 2.1; SalI--2.4, 2.1, 1.6. Recommendation for verification: StuI--4.84, 1.30; SalI--2.4, 2.18, 1.56; PstI--6.14; NotI--6.14.

High level of expression in E. coli, cos and other mammalian cultured cell lines.
- personal communication

The pGFLEX expression system allows production of target proteins fused to either the N or C terminus of the GST pi protein and provides rapid purification of target proteins as either GST fusions or native proteins after cleavage with thrombin.
- Gene 193: 229-237, 1997

History

Depositors
WE Fahl, H Manoharan
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.
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

Manoharan HT, et al. High-level production and purification of biologically active proteins from bacterial and mammanlian cells using the tandem pGFLEX expression system. Gene 193: 229-237, 1997. PubMed: 9256081

Herbert Manoharan, personal communication

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