Addgene - pLKO.1 - TRC cloning vector Plasmid Data

Gene/insert name:		stuffer
Insert size (bp):		1900
Vector backbone:		pLKO.1 (Search Vector Database)
Type of vector:		Mammalian expression,Lentiviral,RNAi
Backbone size (bp):		7032
Cloning site 5':		AgeI
Site destroyed during cloning:		No
Cloning site 3':		EcoRI
Site destroyed during cloning:		No
5' Sequencing primer:		LKO.1 5' (List of Sequencing Primers)
Bacteria resistance:		Ampicillin
High or low copy:		High Copy
Grow in standard E. coli @ 37C:		Yes
Selectable markers:		Puromycin
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		DH5a
Principal Investigator:		David Root
Additional Documents:		Addgene's pLKO.1 protocol (HTML)
Terms and Licenses:		MTA

Comments: This is the recommended vector for cloning and expressing new shRNA sequences. This plasmid is being used by The RNAi Consortium to produce their shRNA library (http://www.broad.mit.edu/genome_bio/trc/). The 1.9kb stuffer can be released with AgeI and EcoRI and replaced with your shRNA sequence of choice.

The link to the author's map shows the key features of this vector before the 1.9kb stuffer was cloned in. The link to the sequence shows the entire sequence with the stuffer sequence in capital letters.

Also, see Addgene's pLKO.1 protocol (http://www.addgene.org/plko) on how to use the pLKO.1 vector.

Plasmid grows more slowly than standard plasmids.

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

Click on map to enlarge

Selected features

ORF frame 3

530 - 8817

ORF frame 1

289 - 1128

ORF frame 1

1179 - 334

puro(variant)

529 - 1128

cPPT

1239 - 1254

U3PPT

1239 - 1260

delta_U3

1256 - 1308

HIV-1_5_LTR

1309 - 1489

truncHIV-1_3_LTR

1309 - 1489

SV40_PA_terminator

1564 - 1683

EBV_rev_primer

1652 - 1671

SV40_origin

1722 - 1799

SV40pro_F_primer

1784 - 1803

T7_promoter

1896 - 1878

M13_forward20_primer

1922 - 1906

M13_pUC_fwd_primer

1937 - 1915

lacZ_a

1915 - 2063

f1_origin

2081 - 2387

AmpR_promoter

2580 - 2608

ORF frame 1

2650 - 3510

Ampicillin

2650 - 3510

pBR322_origin

3665 - 4284

lac_promoter

4593 - 4622

M13_pUC_rev_primer

4636 - 4658

M13_reverse_primer

4657 - 4675

T3_promoter

4692 - 4711

HIV-1_5_LTR

4965 - 5145

truncHIV-1_3_LTR

4965 - 5145

HIV-1_psi_pack

5256 - 5300

RRE

5810 - 6043

ORF frame 3

5688 - 6545

hU6_promoter

6579 - 6811

LKO1_5_primer

6741 - 6760

cPPT

8736 - 8751

Unique restriction sites

SpeI		253
BamHI		510
KpnI		1189
NcoI		1763
NruI		5344
NdeI		6748
AgeI		6816
BstBI		7427
EcoRI		8691

Article: A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Moffat J et al. (Cell. 2006 Mar 24. 124(6):1283-98. 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 10878" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.