Vector: pVALIUM10

Name: pVALIUM10
Description: Drosophila expression vector for high expression of hairpin insert. loxP, and attB recombination sites. Detailed procedure for small scale cloning can be found on the TRiP website: http://flyrnai.org/TRiP-HOME.html Hsp70 promoter with ftz intron and gypsy insulator. 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):: 11143
Properties: drosophila in vitro and in vivo expression
Vector Map: VALIUM10_map_seq.pdf
Vector Sequence: pVALIUM10.txt
Comments: To amplify this vector you will need to use One Shot ccdB Survival 2 T1R Competent Cells(Invitrogen Cat. No. A10460), or other gateway competent cells. 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 618 645
selectable marker vermilion 664 2543
GATEWAY 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 5' TCA ACT GCA ACT ACT GA 3' hsp70(+60)f Recommended for insert verification 3894 3910
Cloning Marker attR1 A Gateway® cassette containing attR recombination sites flanking a ccdB gene and a chloramphenicol-resistance gene 3994 5705
bacterial selection antibiotic CmR 4230 4887
Cloning Marker ccdB toxin 5232 5537
intron ftz Involved in DNA binding. Also known as CG2047-PA,Fushi-Tarazu, Ultra-abdominal-like, ftz-PA, fushi tarazu 5725 5871
Cloning Marker attR2 A Gateway® cassette containing attR recombination sites flanking a ccdB gene and a chloramphenicol-resistance gene 5882 7586
Cloning Marker ccDB #2 6046 6352
bacterial selection antibiotic CmR #2 6696 7353
intron ftz #2 Involved in DNA binding. Also known as CG2047-PA,Fushi-Tarazu, Ultra-abdominal-like, ftz-PA, fushi tarazu 7633 7779
polyA site SV40 7803 8502
repressor binding site Gypsy Insulator #2 8509 8939
primer site T3 8956 8975
primer site M13 reverse 8993 9013
bacterial operon LacO 9018 9040
bacterial origin ColE1 9381 10063
bacterial selection antibiotic AmpR 10161 10820

Authors:

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

Publications:

PMID Title
19487563 A Drosophila resource of transgenic RNAi lines for neurogenetics.

Parent Vector:

Name Comments