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

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

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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 Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1
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

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