Addgene - miR-124-5 promoter GFP Plasmid Data

Plasmid 20792: miR-124-5 promoter GFP

Gene/insert name:
mir-124
Alt name:
miR124
Insert size:
5600
Species:
D. rerio (zebrafish)
Entrez Gene:
mir124-5 (dre-mir-124-5)
Fusion protein or tag:
GFP
Terminal:
C terminal on backbone
Vector backbone:
pBSII SK+ modified to introduce GFP and more cloning sites
(Search Vector Database)
Backbone manufacturer:
Bartel Lab
Vector type:
zebrafish expression
Cloning site 5':
SmaI
Site destroyed during cloning:
Yes
Cloning site 3':
SalI
Site destroyed during cloning:
Unknown
5' sequencing primer:
TGCTTTGAGGCACGGTTATGGTCCC List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
Unknown
Sequence: View sequences (2)
Map:
View map
Principal Investigator:
David Bartel
Terms and Licenses:
MTA

Comments:

5′-RACE (GenRacer kit, Invitrogen) identified a transcriptional start of the mir-124-5 primary transcript, which is located 1.1 kb from the mir-124-5 hairpin. A 5.6-kb fragment upstream of the mir-124-5 transcriptional start was amplified from genomic DNA using primers 1 and 2 (5'-TGCTTTGAGGCACGGTTATGGTCCC-3' and 5'-CTCGGGCAGCGGCTTTTTGGTCCTG-3'). The promoter fragment was fused to a GFP reporter gene. The miRNA promoter fusion was flanked by I-SceI meganuclease recognition sites to increase the efficiency of transgenensis.

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

Article: Coherent but overlapping expression of microRNAs and their targets during vertebrate development. Shkumatava et al (Genes Dev. 2009 Feb 15. 23(4):466-81. 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 20792" in your Materials and Methods section.