Vector: pWALIUM10

Number of plasmids with this backbone: 1
Name: pWALIUM10
Description: Drosophila expression vector for high expression of hairpin insert. Hsp70 promoter with 5xUAS, two gypsy insulators, and two ftz introns. AmpR in bacteria; white marker in drosophila. loxP and GATEWAY recombination sites. Detailed procedure for small scale cloning can be found on the TRiP website: http://flyrnai.org/TRiP-HOME.html Vector may recombine in bacteria and should be stored as DNA.
Synonyms:
Type: bacterial plasmid
Form: dsDNA
Size (bp):: 13426
Properties: drosophila in vitro and in vivo expression
Vector Map: WALIUM10_map_seq.pdf
Vector Sequence: pWALIUM10.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 619 645
selectable marker white 682 4808
Cloning Site attB1 4851 5218
repressor binding site Gypsy Insulator 5230 5660
Cloning Site LoxP 5678 5711
enhancer 5xUAS UAS (Upstream Activation Sequence), a short section of the promoter region, to which Gal4 specifically binds to activate gene transcription 5718 5827
Cloning Site LoxP #2 5834 5867
enhancer 5xUAS #2 UAS (Upstream Activation Sequence), a short section of the promoter region, to which Gal4 specifically binds to activate gene transcription 5874 5983
promoter Hsp70 70 kilodalton heat shock protein 5990 6249
primer site pUAST_Forward tcaactgcaactactga hsp70(+60)f Recommended for insert verification 6177 6193
Cloning Marker attR1 A Gateway® cassette containing attR recombination sites flanking a ccdB gene and a chloramphenicol-resistance gene 6277 7988
bacterial selection antibiotic CmR 6514 7170
toxin ccdB 7515 7820
intron ftz Involved in DNA binding. Also known as CG2047-PA,Fushi-Tarazu, Ultra-abdominal-like, ftz-PA, fushi tarazu 8008 8154
Cloning Marker attR2 A Gateway® cassette containing attR recombination sites flanking a ccdB gene and a chloramphenicol-resistance gene 8165 9869
toxin ccDB #2 8330 8635
bacterial selection antibiotic CmR #2 8980 9636
intron ftz #2 Involved in DNA binding. Also known as CG2047-PA,Fushi-Tarazu, Ultra-abdominal-like, ftz-PA, fushi tarazu 9916 10062
poly-A signal SV40 10086 10785
ribosome binding site Gypsy Insulator #2 10792 11222
primer site T3 11239 11258
primer site M13 reverse 11276 11296
bacterial operon LacO 11302 11324
bacterial origin ColE1 11664 12346
bacterial selection antibiotic AmpR 12444 13103

Authors:

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

Publications:

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

Parent Vector:

Name Comments