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pTRE-Myc Vector Information
GenBank Accession #: Submission in progress.
EcoR I BamH I
CAA GCT TGG TCG ACC GAG ATC TCT CGA GGT ACC GCG GCC GCT CGA CGA TAT CTC TAG A Map and Multiple Cloning Site (MCS) of pTRE-Myc Vector. Unique restriction sites are in bold.
pTRE-Myc is a tetracycline response plasmid that can be used to tag a gene of interest with a c-Myc
epitope and then to induce expression of the tagged protein in Tet-OnTM, Tet-OffTM, RevTet-OnTM, or
RevTet-OffTM Cell Lines. The Tet Expression Systems and Cell Lines give researchers ready access
to the tetracycline-regulated expression systems described by Gossen & Bujard (1; Tet-Off) and
Gossen et al. (2; Tet-On). The pTRE-Myc Vector contains an MCS downstream of the Tet-
responsive P
promoter. cDNAs or genes inserted in the MCS will be responsive to either the tTA or rtTA regulatory protein when expressed by a separate construct in the Tet-Off or Tet-Onsystems, respectively. The P promoter contains the Tet-responsive element (TRE), which consists of seven copies of the 42-bp tet operator sequence (tetO). The TRE element is justupstream of the minimal CMV promoter (P ), which lacks the enhancer that is part of the complete CMV promoter in the pTet plasmids. Consequently, P binding of TetR or rTetR to the tetO sequences. In addition to these elements, pTRE-Myc includesa Kozak consensus ribosome binding site (3), an ATG initiation codon, and the Myc epitope directlyupstream of the MCS; thus, these elements should not be included in the cloned insert. cDNAsinserted in-frame will be expressed as N-terminal Myc-tagged fusions, permitting detection usingCLONTECH’s c-Myc Monoclonal Antibody (#3800-1). To permit selection of stable transfectants,this plasmid should be cotransfected with the pTK-Hyg Vector (not included).
The pTRE-Myc-Luc Control Vector, packaged with the pTRE-Myc Vector, contains an additional1,660 bp encoding firefly luciferase inserted into the Hind III site of the MCS. This vector can be usedas a reporter of induction efficiency using standard luciferase detection reagents. It is not intendedas a cloning vector. In Western blots, myc-luciferase can be detected as a 68-kDa protein.
Vector Information
Location of features:
Location of seven tetO 18-mers: 15–33; 57–75; 99–117; 141–159; 183–201; 225–243 & 257–275 • Fragment containing β-globin poly A signal: 638–1787• Fragment containing Col E1 origin of replication: 1978–2632 • Ampicillin resistance gene (β-lactamase): 3709–2779 Propagation in E. coli:
• Suitable host strains: DH5α and other general purpose strains.
• Selectable marker: plasmid confers resistance to ampicillin (100 µg/ml) to E. coli hosts.
• E. coli replication origin: Col E1 References:
1. Gossen, M. & Bujard, H. (1992) Proc. Natl. Acad. Sci USA 89:5547–5551.
2. Gossen, M., et al. (1995) Science 268:1766–1769.
3. Kozak, M. (1987) Nucleic Acids Res. 15:8125–8148.
Notice to Purchaser
The attached sequence file has been compiled from information in the sequence databases, published literature, and other sources, together
with partial sequences obtained by CLONTECH. This vector has not been completely sequenced.
This product is intended to be used for research purposes only. It is not to be used for drug or diagnostic purposes nor is it intended for human use.
CLONTECH products may not be resold, modified for resale, or used to manufacture commercial products without written approval of CLONTECH.
1999, CLONTECH Laboratories, Inc.
CLONTECH Laboratories, Inc.
Protocol # PT3398-5
Version # PR9X162


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