Addgene - pLL3.7 Plasmid Data

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Luk Van Parijs Lab Plasmids

Mammalian RNAi Tools

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pLKO.1 - TRC cloning vector

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Plasmid 11795: pLL3.7

Gene/insert name:		none
Insert size (bp):		Unknown
Vector backbone:		pLL3.7 (Search Vector Database)
Type of vector:		Mammalian expression,Lentiviral,RNAi,Cre/Lox
Backbone size (bp):		7650
Cloning site 5':		HpaI
Site destroyed during cloning:		No
Cloning site 3':		XhoI
Site destroyed during cloning:		No
5' Sequencing primer:		U6 (5'-cagtgcaggggaaagaatagtaga c-3') (List of Sequencing Primers)
Bacteria resistance:		Ampicillin
High or low copy:		High Copy
Grow in standard E. coli @ 37C:		Yes
Please specify bacterial strain for growth and growth condition:		Although this plasmid grows in standard bacteria, to reduce the chance of recombination we recommend that you grow the plasmid at 30oC in Stbl3 cells from Invitrogen.
Selectable markers:		GFP
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		DH5a
Principal Investigator:		Luk Van Parijs
Additional Documents:		pLL3.7 shRNA cloning (PDF)
Terms and Licenses:		MTA

Comments: Lentiviral vector that expresses shRNA under the mouse U6 promoter. A CMV-EGFP reporter cassette is included in the vector to monitor expression.

For protocols regarding the use of pLL3.7, visit: (http://web.mit.edu/jacks-lab/protocols/rnairesources.htm).

Note: A single base pair deletion at position 11 of the U6 promoter in this plasmid does not impair the efficacy of this reagent.

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

1686 - 1919

ORF frame 1

1564 - 2451

cPPT

2450 - 2465

pBluescriptKS_primer

2951 - 2967

loxP

3010 - 3043

CAG_enhancer

3163 - 3450

CMV_immearly_promoter

3108 - 3660

CMV_fwd_primer

3617 - 3637

CMV_promoter

3618 - 3687

EGFP_N_primer

3777 - 3756

EGFP

3711 - 4427

ORF frame 3

3711 - 4430

ORF frame 3

4466 - 3696

EGFP_C_primer

4364 - 4385

loxP

4450 - 4483

WPRE

4541 - 5128

pBluescriptKS_primer

5147 - 5131

ORF frame 1

4642 - 5682

cPPT

5319 - 5334

U3PPT

5319 - 5340

HIV-1_5_LTR

5656 - 5836

truncHIV-1_3_LTR

5656 - 5836

pBR322_origin

6502 - 5883

ORF frame 3

7517 - 6657

Ampicillin

7517 - 6657

AmpR_promoter

7587 - 7559

Unique restriction sites

SpeI		252
NarI		1019
PstI		2420
ApaI		2612
XbaI		2616
HpaI		2935
XhoI		2950
NotI		3065
NheI		3689
AgeI		3698
EcoRI		4431
SacII		5042
KpnI		5269
PvuII		5296
FspI		6952

Article: A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Rubinson DA et al. (Nat Genet. 2003 Mar . 33(3):401-6. 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 11795" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.