pTK-HSV-BP2 plasmid in Escherichia coli (ATCC® 99530)

Organism: Homo sapiens, human  /  Clone Type: Clone  /  Depositors: JL Goldstein

Permits and Restrictions

View Permits

Designations pTK-HSV-BP2 plasmid in Escherichia coli
GenBank Number

U02031

Species Homo sapiens, human
Depositors JL Goldstein
Applications
in suitable host, produces protein sterol regulatory element binding transcription factor 2 sterol regulatory element-binding protein 2
Vector
Name of vector: pTK-HSV
Intact vector size: 5.8
Type of vector: phagemid
Vector end: BspDI
Vector end: XbaI
Cloning sites: BspDI XbaI
Construction: pcDNA3
Host range: mammalian cells; Escherichia coli


Features (with orientation and position when available):
replicon: pMB1
marker(s): neoR, ←
replicon: f1, ←
terminator: bGH polyadenylation
restriction site: XbaI
restriction site: BspDI
epitope tag: HSV
promoter: HSV TK, ←
marker(s): ampR, →
Insert Insert size (kb): 4.2
Gene: sterol regulatory element binding transcription factor 2, SREBP2
DNA: cDNA
Source: human
Genbank accession : U02031
Nucleotides: Nucleotides 1-4093 of the insert correspond to nucleotides 156-4249 of U02031
Insert Size (kb) 4.2
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Shipping Information Freeze dried E. coli containing the plasmid
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.
References

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