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Detailed Vector Information: pLIFE-miRNA

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Vector Name: pLIFE-miRNA
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Synonyms: None

pLIFE-miRNA

Sequencing Primer: Forward:  M13F
Reverse:  M13R
Description: Mammalian expression vector for expressing miRNAs; also contains red fluorescent protein, gateway cassette located within an intron of RFP 3'UTR, for ncRNA expression; ampicillin resistance in bacteria; Gateway cloning with L2R3 Gateway Cassette
Comments: Growth before gateway recombination requires DB3.1 E. coli cells.
Size (bp): 6500
Empty Vector: EvNO00601504
Parent Vector: None
Properties: Gateway, acceptor (destination), fluorescent marker, mammalian expression, recombinational cloning
Author Name: Center for Personalized Diagnostics
Arizona State University
Marco Mangone
Justin Wolter
Publications: PMID: 25074381
Title: 3pLIFE: a functional assay to detect miRNA targets in high-throughput

Vector Map:         Vector Sequence:
Sequence:

 



Vector Features:

Type Name Description Start Position End Position
3pUTR beta globin rabbit beta-globin polyadenylation signal 2484 3035
bacterial origin ColE ColE1 origin 4143 4731
fluorescent marker DsRed mammalian codon-optimized tetrameric variant of DsRed fluorescent protein 7 684
primer M13 forward M13 forward sequencing primer 2412 2428
promoter CMV CMV promoter 5925 6432
selectable marker CmR Chloramphenicol resistance (part of theL2R3 Gateway Cassette); removed in plasmids containing a miRNA 1516 2172
selectable marker AmpR ampicillin resistance 4902 5867
splice acceptor site 3' splice acceptor 3'splice site 2435 2483
splice donor site 5' splice donor 5' splice site 693 701