Addgene - pLKO.3G Plasmid Data

Gene/insert name:		none
Insert size (bp):		Unknown
Vector backbone:		pLKO.3G (Search Vector Database)
Type of vector:		Mammalian expression,Lentiviral,RNAi
Backbone size (bp):		8174
Cloning site 5':		EcoRI
Site destroyed during cloning:		No
Cloning site 3':		PacI
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:		eGFP
If you did not originally clone this gene, from whom and where did you receive the plasmid used to derive this plasmid:		Derived from the pLKO.1 plasmid from The RNAi Consortium (TRC)
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		Stbl3
Principal Investigator:		Christophe Benoist, Diane Mathis
Additional Documents:		Addgene's pLKO.1 protocol (HTML)
Terms and Licenses:		MTA

Comments: A variant of pLKO.1 containing an eGFP marker rather than puromycin. Please note that the AgeI site is NOT unique for this vector, so shRNAs should be designed for cloning with EcoRI and PacI instead. Forward oligo: 5'-AATT—21bp sense—CTCGAG—21bp antisense—TTTTTTTAT-3'. Reverse oligo: 5'-AAAAAAA—21bp sense—CTCGAG—21bp antisense-3'.

pLKO.3G was cloned by excising PAC-GFP from pLKO.3pgw and replacing it with eGFP from another vector using the enzymes BamHI and BsRGI.

See http://citdh.org/Lentivectors.html for more information.

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

Click on map to enlarge

Selected features

cPPT

168 - 183

EGFP_N_primer

999 - 978

ORF frame 3

693 - 1652

EGFP

933 - 1649

EGFP_C_primer

1586 - 1607

WPRE

1673 - 2260

ORF frame 1

1774 - 2544

pBluescriptKS_primer

2279 - 2263

cPPT

2448 - 2463

U3PPT

2448 - 2469

delta_U3

2465 - 2517

HIV-1_5_LTR

2518 - 2698

truncHIV-1_3_LTR

2518 - 2698

SV40_PA_terminator

2773 - 2892

EBV_rev_primer

2861 - 2880

SV40_origin

2931 - 3008

SV40pro_F_primer

2993 - 3012

T7_promoter

3105 - 3087

M13_forward20_primer

3131 - 3115

M13_pUC_fwd_primer

3146 - 3124

lacZ_a

3124 - 3272

f1_origin

3290 - 3596

AmpR_promoter

3789 - 3817

ORF frame 1

3859 - 4719

Ampicillin

3859 - 4719

pBR322_origin

4874 - 5493

lac_promoter

5802 - 5831

M13_pUC_rev_primer

5845 - 5867

M13_reverse_primer

5866 - 5884

T3_promoter

5901 - 5920

HIV-1_5_LTR

6174 - 6354

truncHIV-1_3_LTR

6174 - 6354

HIV-1_psi_pack

6465 - 6509

RRE

7019 - 7252

ORF frame 3

6897 - 7754

hU6_promoter

7983 - 65

LKO1_5_primer

8169 - 14

Unique restriction sites

NdeI		2
EcoRI		94
MscI		723
BamHI		914
EcoRV		1655
AatII		2124
KpnI		2398
StuI		3064
NruI		6553
NotI		6864

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

Also, please include the text "Addgene plasmid 14748" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.