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Browse > Patrick Aebischer > Szulc et al > pLVUT-tTR-KRAB

 

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This is commonly
requested with
pLVCT-tTR-KRAB
pLVTHM
pLVPT-tTR-KRAB
pMD2.G
psPAX2
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Plasmid 11651: pLVUT-tTR-KRAB
Gene/insert name: hUbiquitin C, GFP, tTR-KRAB, Tet-on
Insert size (bp): Unknown
Vector backbone: None  (Search Vector Database)
Type of vector: Mammalian expression,Lentiviral
Backbone size (bp): 11480
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

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.

Note: There is another EcoRI site at 6235 that is not depicted in the author's map and sequence.

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

Click on map to enlarge

Selected features
CAG_enhancer 318 - 605
CMV_immearly_promoter 239 - 815
CMV_fwd_primer 772 - 792
HIV-1_5_LTR 835 - 1015
truncHIV-1_3_LTR 835 - 1015
HIV-1_psi_pack 1126 - 1170
RRE 1680 - 1913
Orf frame 1 1558 - 2445
cPPT 2444 - 2459
hUbC_promoter 2617 - 3833
EGFP_N_primer 3926 - 3905
EGFP 3860 - 4576
Orf frame 2 3860 - 4579
EGFP_C_primer 4513 - 4534
IRES 4614 - 5117
TetR 5195 - 5794
Orf frame 2 5180 - 6187
WPRE 6256 - 6843
Orf frame 2 6344 - 6994
pBluescriptKS_primer 6862 - 6846
cPPT 6916 - 6931
U3PPT 6916 - 6937
TRE 6968 - 7255
HIV-1_5_LTR 7287 - 7467
truncHIV-1_3_LTR 7287 - 7467
BGH_rev_primer 7508 - 7491
bGH_PA_terminator 7494 - 7721
f1_origin 7784 - 8090
pBABE_3_primer 8224 - 8204
SV40_enhancer 8425 - 8210
SV40_promoter 8222 - 8490
SV40_origin 8389 - 8466
SV40pro_F_primer 8451 - 8470
EM7_promoter 8584 - 8651
bleo 8652 - 9023
sh_ble 8652 - 9026
SV40_PA_terminator 9159 - 9278
EBV_rev_primer 9247 - 9266
M13_reverse_primer 9340 - 9322
M13_pUC_rev_primer 9361 - 9339
lac_promoter 9404 - 9375
pBR322_origin 10332 - 9713
Orf frame 2 11347 - 10487
Ampicillin 11347 - 10487
AmpR_promoter 11417 - 11389
Unique restriction sites

PstI 2414
BamHI 3835
AgeI 3847
EcoRI 4596
ApaI 4720
BstBI 6232
MscI 8655
FspI 10782

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 11651" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.

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