Vector: pVALIUM20

Name: pVALIUM20
Description: Drosophila expression vector for high expression of hairpin insert. loxP, and GATEWAY recombination sites, NheI and EcoRI method to clone shmiR. Detailed procedure for small scale cloning can be found on the TRiP website: http://flyrnai.org/TRiP-HOME.html Hsp70 promoter with ftz intron, two gypsy insulators, 5xUAS and vermilion selection in drosophila. AmpR in bacteria; vermilion marker in drosophila. Vector may recombine in bacteria and should be stored as DNA.
Synonyms:
Type: bacterial plasmid
Form: dsDNA
Size (bp):: 8190
Properties: drosophila in vitro and in vivo expression
Vector Map: VALIUM20_map_seq.pdf
Vector Sequence: pVALIUM20.txt
Comments: Revised 4/25/2012

Features:

Type Name Description Start Position End Position
bacterial origin F1 origin 138 444
bacterial operon LacZ alpha 463 531
primer site M13 forward 602 619
promoter T7 619 645
selectable marker vermilion 664 2543
Cloning Site attB1 2568 2935
repressor binding site Gypsy Insulator 2947 3377
Cloning Site LoxP 3395 3428
enhancer 5xUAS UAS (Upstream Activation Sequence), a short section of the promoter region, to which Gal4 specifically binds to activate gene transcription 3435 3544
Cloning Site LoxP #2 3551 3584
enhancer 5xUAS #2 UAS (Upstream Activation Sequence), a short section of the promoter region, to which Gal4 specifically binds to activate gene transcription 3591 3700
promoter Hsp70 70 kilodalton heat shock protein 3707 3966
primer site pUAST_Forward TCAACTGCAACTACTGA hsp70(+60)f Recommended for insert verification 3894 3910
restriction site Nhe I 4018 4023
primer site M13 Forward #2 4049 4066
restriction site EcoRI 4635 4640
intron ftz Involved in DNA binding. Also known as CG2047-PA,Fushi-Tarazu, Ultra-abdominal-like, ftz-PA, fushi tarazu 4747 4893
viral gene SV40 poly A 4917 5616
repressor binding site Gypsy Insulator #2 5623 6053
primer site T3 6069 6088
primer site M13 reverse 6106 6126
bacterial operon LacO 6132 6154
bacterial origin ColE1 6494 7176
bacterial selection antibiotic AmpR 7275 7934

Authors:

Author Name Author Type Creation Date
Norbert Perrimon Academic researcher, vector PI
Jian-Quan Ni Academic researcher, vector creator

Publications:

PMID Title
21460824 A genome-scale shRNA resource for transgenic RNAi in Drosophila.

Parent Vector:

Name Comments