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
