Addgene - pSico Plasmid Data

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Plasmid 11578: pSico

Gene/insert name:		none
Insert size (bp):		Unknown
Relevant mutations/deletions:		EGFP is expressed from this plasmid as a marker, but it is not a fusion protein. Cre causes EGFP to be recombined out of the construct, activating shRNA expression.
Vector backbone:		pSico (Search Vector Database)
Type of vector:		Mammalian expression,Lentiviral,RNAi,Cre/Lox
Backbone size (bp):		7566
Cloning site 5':		HpaI
Site destroyed during cloning:		No
Cloning site 3':		XhoI
Site destroyed during cloning:		No
5' Sequencing primer:		EGFP-C (List of Sequencing Primers)
Bacteria resistance:		Ampicillin
High or low copy:		High Copy
Grow in standard E. coli @ 37C:		Yes
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		DH5a
Principal Investigator:		Tyler Jacks
Additional Documents:		pSico shRNA cloning (PDF)
Terms and Licenses:		MTA

Comments: pSico allows for conditional (Cre-Lox), stable expression of shRNAs for RNA interference in cells and transgenic mice. Addition of Cre TURNS ON shRNA expression.

The shRNA coding oligos have to be cloned into the HpaI and XhoI restriction sites. Oligo design information can be found on the author's map or on the Jacks Lab website (http://web.mit.edu/ccr/labs/jacks/protocols/pSico.html).

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

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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

loxP

2925 - 2892

loxP

2892 - 2925

CAG_enhancer

3029 - 3316

CMV_immearly_promoter

2974 - 3526

CMV_fwd_primer

3483 - 3503

CMV_promoter

3484 - 3553

EGFP_N_primer

3643 - 3622

Orf frame 1

4305 - 3562

EGFP

3577 - 4293

Orf frame 1

3577 - 4296

EGFP_C_primer

4230 - 4251

loxP

4337 - 4304

loxP

4304 - 4337

pBluescriptKS_primer

4363 - 4379

WPRE

4457 - 5044

pBluescriptKS_primer

5063 - 5047

Orf frame 1

4558 - 5598

cPPT

5235 - 5250

U3PPT

5235 - 5256

HIV-1_5_LTR

5572 - 5752

truncHIV-1_3_LTR

5572 - 5752

pBR322_origin

6418 - 5799

Orf frame 3

7433 - 6573

Ampicillin

7433 - 6573

AmpR_promoter

7503 - 7475

Unique restriction sites

SpeI		252
NarI		1019
PstI		2420
ApaI		2612
XbaI		2616
EcoRI		2941
NheI		3555
AgeI		3564
NotI		4297
HpaI		4347
XhoI		4362
SacII		4958
KpnI		5185
PvuII		5212
FspI		6868

Article: Cre-lox-regulated conditional RNA interference from transgenes. Ventura A et al. (Proc Natl Acad Sci U S A. 2004 Jul 13. 101(28):10380-5. 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 11578" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.