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pGEX-5G/LIC

77274

Clone type
Vector
Applications
Molecular biology
Product format
Freeze-dried
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
expression vector
vector permitting construction of fusion proteins

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): SacII--5.1; PstI--5.1; SalI--5.1; PstI/BamHI--4.0, 0.98.
Vector is covered by a U.S. patent application and is distributed only for non-commercial use.
The carrier protein can be removed by thrombin proteolysis.
Preparation of the vector for cloning includes linearization with SacII, gel purification of the linearized vector, and treatment with T4 DNA polymerase in the presence of dATP.
Target sequences for cloning are prepared by PCR and do not require restriction enzyme digestion.
Annealing the vector and amplification product forms a gapped duplex molecule that can be used directly to transform bacteria without ligation.
Ligation-independent expression vector for constructing fusion proteins with a carrier polypeptide (glutathione S-transferase) which allows single-step affinity purification.
The forward primer should contain 15 nt complementary to nt 5' to the SacII site of the vector [(5'-3')(GGCCTGGTTCCGCGG)] followed by 12-15 nt corresponding to the target sequence (nt encoding the N-terminal 4-5 aa of the protein).
The reverse primer should contain 14 nt complementary to nt 3' to the SacII site of the vector [(5'-3') (CTGCGCCTCGCTCC)] followed by 12 nt complementary to the 3' end of the target sequence.
Both primers should contain a dAMP residue near the sequence complementary to the vector to terminate the exonucleolytic activity of the subsequent T4 DNA polymerase treatment.
Design of PCR primers permits cloning in any reading frame.
The order of the major features in this plasmid is: pMB1 ori - lacIq - lacZ' - tac - GST/SalI/SacII/thrombin cleavage site/BamHI - ampR.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
4.967999935150147
Vector name
pGEX-5G/LIC (plasmid)
Type of vector
plasmid
Construction
pGEX-2T
Markers
ampR
Promoters
tac
Replicon
pMB1
Repressor gene
lacIq

Handling information

History

Depositors
RS Haun
Cross references
GenBank M97937

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

Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67: 31-40, 1988. PubMed: 3047011

Haun RS, Moss J. Ligation-independent cloning of glutathione S-transferase fusion genes for expression in Escherichia coli. Gene 112: 37-43, 1992. PubMed: 1339364

Randy S Haun, personal communication

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