pHSREM4

One of a series (ATCC 37642-37645) of plasmid vectors for expressing genes under the control of the Drosophila hsp70 promoter & terminator. The transcription module is flanked by restriction sites (facilitating its removal), T7 and T3 promoters (for in vitro RNA synthesis), & primers for Sanger sequencing. There is also an M13 origin of replication. A KpnI/SacI double digest releases the hsp70 transcription module, for subsequent ligation into other plasmids such as ones containing Drosophila P elements. KpnI BglII NotI XhoI – promoter – MCS – terminator – SpeI XbaI NotI EagI BstXI SacII SacI. Restriction digests of the clone give the following sizes (kb): PstI–3.3; HindIII–3.3; EcoRI–3.3. (ATCC staff) The KpnI/SacI fragment of pHSREM1 (ATCC 37642) containing the hsp70 transcription module was ligated into KpnI/SacI digested Bluescript SK-. (personal communication) Major Drosophila hsp70 regulator elements are: 2 heat shock consensus elements (-85 to -72 & -62 to -49),a TATA box (-33 to -26) & transcription initiation site (+1) in the 5′ segment; the termination triplet and polyA site in the 3′ segment. (personal communication) Medium is 1227 LB plus ampicillin. Hosts: E.coli HB101, E.coli.