Addgene - FCK-Halo-GFP Plasmid Data

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Edward Boyden Lab Plasmids

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FCK-ChR2-GFP

pCAGGS-ChR2- Venus

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Plasmid 14750: FCK-Halo-GFP

Gene/insert name:		Mammalian codon-optimized halorhodopsin, from Natronomas pharaonis (N. pharaonis), fused with GFP at the C-terminus
Alternative names:		Halo
		Halo-GFP
		NPhR
Insert size (bp):		1300
Species of gene(s):		N. pharaonis
Relevant mutations/deletions:		N. pharaonis halorhodopsin is mammalian codon-optimized
Fusion proteins or tags:		GFP
Terminal:		C terminal on insert
Vector backbone:		FCK(1.3)GW (Search Vector Database)
Backbone manufacturer:		Pavel Osten
Type of vector:		Mammalian expression,Bacterial expression,Lentiviral
Backbone size (bp):		9500
Cloning site 5':		AgeI
Site destroyed during cloning:		No
Cloning site 3':		EcoRI
Site destroyed during cloning:		No
5' Sequencing primer:		atgactgagaccctcccacccgtg actgaaagcgccgt (List of Sequencing Primers)
Bacteria resistance:		Ampicillin
High or low copy:		Unknown
Grow in standard E. coli @ 37C:		No
Please specify bacterial strain for growth and growth condition:		Use recombinase-free E. coli (e.g., Invitrogen's Stbl3)
If you did not originally clone this gene, from whom and where did you receive the plasmid used to derive this plasmid:		We purchased the gene to be synthesized by Epoch Biolabs (Sugar Land, TX)
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		Stbl3
Principal Investigator:		Edward Boyden
Terms and Licenses:		MTA

Comments: PLEASE CONTACT ED BOYDEN (esb@media.mit.edu) FOR DETAILS ON THIS REAGENT AND FURTHER REAGENTS IN THIS LINE.

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

1680 - 1913

ORF frame 1

1558 - 2445

cPPT

2444 - 2459

ORF frame 1

4425 - 3703

ORF frame 1

3745 - 5511

EGFP_N_primer

4858 - 4837

EGFP

4804 - 5508

EGFP_C_primer

5445 - 5466

WPRE

5538 - 6125

pBluescriptKS_primer

6144 - 6128

ORF frame 1

5626 - 6675

cPPT

6312 - 6327

U3PPT

6312 - 6333

HIV-1_5_LTR

6649 - 6829

truncHIV-1_3_LTR

6649 - 6829

BGH_rev_primer

6872 - 6855

bGH_PA_terminator

6858 - 7085

f1_origin

7148 - 7454

pBABE_3_primer

7588 - 7568

SV40_enhancer

7789 - 7574

SV40_promoter

7586 - 7854

SV40_origin

7753 - 7830

SV40pro_F_primer

7815 - 7834

EM7_promoter

7948 - 8015

bleo

8016 - 8387

sh_ble

8016 - 8390

SV40_PA_terminator

8523 - 8642

EBV_rev_primer

8611 - 8630

M13_reverse_primer

8704 - 8686

M13_pUC_rev_primer

8725 - 8703

lac_promoter

8768 - 8739

pBR322_origin

9696 - 9077

ORF frame 2

10711 - 9851

Ampicillin

10711 - 9851

AmpR_promoter

10781 - 10753

Unique restriction sites

SpeI		252
NdeI		487
NarI		1019
NotI		1525
XbaI		2651
BamHI		3895
AgeI		4782
EcoRI		5512
SacII		6039
XhoI		6140
KpnI		6262
FspI		10146

Article: Multiple-color optical activation, silencing, and desynchronization of neural activity, with single-spike temporal resolution. Han X et al. (PLoS ONE. 2007 . 2():e299. 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 14750" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.