Addgene - pLVPRT-tTR-KRAB Plasmid Data

Gene/insert name:		hPrion, GFP, tTR-KRAB, Tet-on
Insert size (bp):		Unknown
Vector backbone:		None (Search Vector Database)
Type of vector:		Mammalian expression,Lentiviral
Backbone size (bp):		11583
5' Sequencing primer:		See map (List of Sequencing Primers)
Bacteria resistance:		Ampicillin
High or low copy:		High Copy
Grow in standard E. coli @ 37C:		No
Please specify bacterial strain for growth and growth condition:		Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		Stbl3
Principal Investigator:		Patrick Aebischer
Terms and Licenses:		MTA, Tet Systems LULL

Comments: Transgenes can be expressed from any RNA Pol II promoter as part of bicistronic unit comprising the KRAB-based repressor; tetO sequences are inserted into the vector LTR. Tet-on and Tet-off versions rely on repressors that bind in the absence or the presence of doxycycline, respectively. Addition of Pol III promoter-small hairpin RNA cassette allows for drug-controllable RNA interference (Tet-on shRNA).

pLVTHM and packaging plasmid for this system are also available at Addgene (http://www.addgene.org/rnaitools). Please visit Trono lab website (http://tronolab.epfl.ch) to see frequently asked questions on cloning strategies and packaging. You may also visit LentiWeb (http://www.lentiweb.com) for discussion on cloning strategies and protocols.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Click on map to enlarge

Selected features

HIV-1_5_LTR

461 - 641

truncHIV-1_3_LTR

461 - 641

HIV-1_psi_pack

752 - 796

RRE

1306 - 1539

synth_int

2359 - 2634

EGFP_N_primer

2789 - 2768

EGFP

2723 - 3439

ORF frame 2

2723 - 3472

EGFP_C_primer

3376 - 3397

IRES

3523 - 4026

TetR

4104 - 4703

ORF frame 3

4089 - 5096

WPRE

5171 - 5758

cPPT

5829 - 5844

U3PPT

5829 - 5850

TRE

5881 - 6168

HIV-1_5_LTR

6204 - 6384

truncHIV-1_3_LTR

6204 - 6384

Sp6_primer

6427 - 6410

pBRrevBam_primer

6736 - 6717

tet(564-300)

6928 - 6665

pGEX_3_primer

7069 - 7091

AmpR_promoter

7250 - 7278

ORF frame 3

7320 - 8180

Ampicillin

7320 - 8180

pBR322_origin

8335 - 8954

pBABE_3_primer

9200 - 9180

SV40_enhancer

9401 - 9186

SV40_promoter

9198 - 9466

SV40_origin

9365 - 9442

SV40_promoter

9318 - 9520

SV40pro_F_primer

9427 - 9446

SV40_int

10647 - 10662

SV40_3_splice

10668 - 10715

SV40_PA_terminator

11291 - 11422

EBV_rev_primer

11379 - 11398

Unique restriction sites

NotI		1151
SmaI		3503
PstI		3511
SpeI		5155
XbaI		6388
FspI		7884
HpaI		11287

Article: A versatile tool for conditional gene expression and knockdown. Szulc J et al. (Nat Methods. 2006 Feb . 3(2):109-16. Pubmed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication.

Also, please include the text "Addgene plasmid 11648" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.