10XSTAT92E luciferase | Addgene 37393 product information

This webpage contains legacy information. The product is either no longer available from the supplier or has been delisted at Labome.

product summary

company name :

Addgene

product type :

cDNA

product name :

10XSTAT92E luciferase

catalog :

37393

citations: 2

Reference
Amarnath S, Stevens L, Stein D. Reconstitution of Torso signaling in cultured cells suggests a role for both Trunk and Torso-like in receptor activation. Development. 2017;144:677-686 pubmed publisher
Baeg G, Zhou R, Perrimon N. Genome-wide RNAi analysis of JAK/STAT signaling components in Drosophila. Genes Dev. 2005;19:1861-70 pubmed

product information

Catalog Number :

37393

Product Name :

10XSTAT92E luciferase

article :

doi	10.1101/gad.1320705
id	5481
pubmed_id	16055650

bacterial resistance :

Ampicillin

cloning :

backbone

pUAST

backbone_mutation

backbone_origin

backbone_size

8904

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Insect Expression

Luciferase

Other

JAK/STAT reporter construct

growth notes :

Perrimon Lab plasmid #446 A 441-bp genomic fragment in the enhancer of SOCS36E containing two potential STAT92E-binding sites was amplified by PCR, using five different sets of oligos: (1) CTGCAGGAACCACTCAGAGTGCCTGCGTGT (PstI), GAATTCATACAAAACTGTCTTAGGTGTTTA (EcoRI); (2) CTGCAGGAACCACTCAGAGTGCCTGCGTGT (PstI), CTGCAGATACAAAACTGTCTTAGGTGTTTA (PstI); (3) GAATTCGAACCACTCAGAGTGCCTGCGTGT (EcoRI), GAATTCATACAAAACTGTCTTAGGTGTTTA (EcoRI); (4) AGATCTGAACCACTCAGAGTGCCTGCGTGT (BglII), AGATCTATACAAAACTGTCTTAGGTGTTTA (BglII); (5) GCGGCCGCGAACCACTCAGAGTGCCTGCGTGT (NotI), GCGGCCGCATACAAAACTGTCTTAGGTGTTTA (NotI). Each amplified genomic fragment containing different restriction enzyme sites was sequentially subcloned into pUAST. The genomic fragment amplified using the first set of oligos was subcloned into the PstI/EcoRI sites of pUAST to generate 2XSTAT92E. The genomic fragment amplified using the second set of oligos was subcloned into the PstI site of 2XSTAT92E to generate 4XSTAT92E. The genomic fragment amplified using the third set of oligos was subcloned into the EcoRI site of 4XSTAT92E to generate 6XSTAT92E. The genomic fragment amplified using the fourth set of oligos was subcloned into the BglII site of 6XSTAT92E to generate 8XSTAT92E. Next, the hsp70 minimal promoter element was amplified from pUAST by PCR using oligos GCGGCCGCAGCGGAGACTCTAGCGAGCG (NotI) and CTCGAGAATTCCCTATTCAGAGTTCT (XhoI). This hsp70 minimal promoter was subcloned into the NotI/XhoI sites of 8XSTAT92E to generate 8XSTAT92E hsp70. Again, the genomic fragment amplified using the fifth set of oligos was subcloned into the NotI site of 8XSTAT92E hsp70 vector to generate 10XSTAT92E hsp70. Finally, an XhoI/XbaI fragment containing the firefly luciferase gene from the pGL3 luciferase vector (Promega) was subcloned into the XhoI/XbaI sites of 10XSTAT92E hsp70 to generate 10XSTAT92E luciferase.

origin :

pi :

alt_names

10X STAT92E binding sites

JAK/STAT reporter construct

SOCS36E

cloning

clone_method	Restriction Enzyme
cloning_site_3	multiple (see below for details)
cloning_site_5	multiple (see below for details)
promoter	Hsp70
sequencing_primer_3
sequencing_primer_5	pUAST-5 (5'-CTGCAACTAAAATCTGCCAAGAAG-3')
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	Dmel_CG15154, CG15154, Dmel CG15154, SOCS, SOCS36E, Socs, anon-EST:Liang-2.2, clone 2.2, dSOCS, dmSocs36E, socs36E, socs60E1
gene	Socs36E
id	35085

genbank_ids

mutation

name

SOCS36E enhancer fragment

shRNA_sequence

size

species

7227

Drosophila melanogaster