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pTK-HSV-BP2 plasmid in Escherichia coli

99530

This construct allows the exogenously expressed SREBP2 to be cleaved in a sterol-regulated fashion after transfection into 293 cells (ATCC CRL-1573). The amino terminus was truncated at amino acid 14 and fused to the HSV tag of the vector.

Name of construct: pTK-HSV-BP2
Size of construct (kb): 10.0
Markers: ampR, neoR
Excise insert: BapDI + XbaI

Depositor: Joseph L. Goldstein, Department of Molecular Genetics, University of Texas Southwestern Medical Center, Callas, TX
Organism
Homo sapiens, human
Clone type
Clone
Applications
Molecular biology
Product format
Freeze-dried
Shipping information
Escherichia coli containing the plasmid
Storage conditions
2°C to 8°C
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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
in suitable host, produces protein sterol regulatory element binding transcription factor 2 sterol regulatory element-binding protein 2

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): BspDI/XbaI--5.8, 4.2; EcoRI--6.8, 3.0; HindIII--9.0; 0.8.
The insert contains the following restriction sites (approximate kb from the 5' end): EcoRV--1.08; HindIII--1.12; BglII--1.70; EcoRI--2.45
This construct allows the exogenously expressed SREBF2 to be cleaved in a sterol-regulated fashion after transfection into 293 cells (ATCC CRL-1573).
The amino terminus was truncated at amino acid 14 and fused to the HSV tag of the vector.
Mycoplasma contamination
Not detected

Vector information

Intact vector size
5.8
Vector name
pTK-HSV
Type of vector
phagemid
Construction
pcDNA3
Host range
mammalian cells
Vector end
BspDI; XbaI
Vector information
epitope tag: HSV
Cloning sites
BspDI; XbaI
Markers
neoR; ampR
Promoters
HSV TK
Replicon
pMB1
Restriction sites
BspDI; XbaI
Terminator
bGH polyadenylation

Insert information

Insert size (kb)
4.2000000000000002
Insert information
Gene: sterol regulatory element binding transcription factor 2, SREBP2
Source: human
Genbank accession : U02031
Nucleotides: Nucleotides 1-4093 of the insert correspond to nucleotides 156-4249 of U02031

Handling information

Host
Escherichia coli HB101 (ATCC 33694)
Medium
Temperature
37°C
Handling procedure
  1. Open vial according to instructions.
  2. Asceptically add 0.3 to 0.4 mL of liquid medium to the freeze-dried pellet and mix well.  Transfer 100 µL to a test tube containing 5 mL LB+ ampicillin (50-100 µg/mL). A loopful of culture can also be streaked on an agar plate of the same. Incubate cultures at 37°C.
  3. Isolate DNA using standard plasmid preparation procedures.
Handling notes
Restriction digests of the clone give the following sizes (kb): BspDI/XbaI--5.8,
4.2; EcoRI--6.8, 3.0; HindIII--9.0; 0.8.
- ATCC staff

The insert contains the following restriction sites (approximate kb from the 5'
end): EcoRV--1.08; HindIII--1.12; BglII--1.70; EcoRI--2.45
- GenBank/EMBL/DDBJ

This construct allows the exogenously expressed SREBF2 to be cleaved in a
sterol-regulated fashion after transfection into 293 cells (ATCC CRL-1573). The
amino terminus was truncated at amino acid 14 and fused to the HSV tag of the
vector.
- personal communication

History

Depositors
JL Goldstein
Cross references
GenBank U02031

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

Hua X, et al. Hairpin orientation of sterol regulatory element-binding protein-2 in cell membranes as determined by protease protection. J. Biol. Chem. 270: 29422-29427, 1995. PubMed: 7493979

Joseph L Goldstein, personal communication

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