pHS24 (ATCC® 37841)

Applications: contains sequence alcohol dehydrogenaseintegrating vectorshuttle vector  /  Depositors: HG Sass

Permits and Restrictions

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Designations pHS24
Depositors HG Sass
Biosafety Level 1
Vector Information
Size (kb): 12.0000000000000000
Vector: pHS24 (plasmid)
Promoters: Promoter hsp82
Construction: pHS17
Marker(s):G418R,ampR,Adh+
Construct size (kb): 12.0
Features: marker(s): ampR, G418R, Adh+
promoter: hsp82
replicon: pMB1
terminator: hsp82
Applications
contains sequence alcohol dehydrogenase
integrating vector
shuttle vector
Comments
Restriction digests of the clone give the following sizes (kb): KpnI--12.5; XbaI--11.0, 0.72, 0.50, 0.37; BamHI/KpnI--7.6, 3.8, 1.3.
Expression is constitutive, but can be increased with heat.
Selection of transgenic Drosophila by G418 has been done with embryos, larvae, and adults. Adults can be reselected by 6% ethanol.
Can be used in Drosophila spp. cell culture transfection assays, and may function in other animal and plant systems because the hsp82 promoter is highly conserved.
P-transposable (integrating) shuttle vector.
Contains hsp82-neo fusion gene which encodes the first 20 aa of D. pseudoobscura hsp82, 5 codons in a linker, fused to the fifth codon of bacterial neomycin phosphotransferase from Tn5, and followed by polyadenylation signals from D. melanogaster hsp82.
Also contains a complete alcohol dehydrogenase coding region from D. melanogaster (3.2 kb XbaI fragment) which includes 0.66 kb of 5' and 0.64 kb of 3' DNA, and has all the cis-acting sequences necessary for expression.
The order of the major components of this construct are: 3' P-element, 5'-Adh-3', 5'-hsp82-neo fusion-hsp82 polyadenylation signal-3', KpnI site, and 5' P-element.
Differs from pHS22 (ATCC 37840) by the removal of hsp82 upstream sequences from approximately -5000 to -868 (where the transcription start point is +1).
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Sass H. P-transposable vectors expressing a constitutive and thermoinducible hsp82-neo fusion gene for Drosophila germline transformation and tissue-culture transfection. Gene 89: 179-186, 1990. PubMed: 2165019

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