Vector IG Sequence Link :
General : plasmid ds-DNA 3400 BP
Functions : (cloning)
Selection : ()
Copy Number :
Hosts : (E.coli HB101)(E.coli)
Suppliers : (ATCC)
Misc.Comments : 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. 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. Restriction digests of the clone give the following sizes (kb): PstI--3.4; HindIII--3.3; EcoRI--3.4; BamHI--3.3. (ATCC staff) BamHI/BglII fragment of pCIII1 with terminator plus several upstream sites of Bluescript MCS ligated into BamHI of pCII38 = pCIII8. KpnI/XhoI region of pCIII8 replaced with synthetic adapter with NotI and BglII sites = pHSREM1. (personal communication) XhoI/HindIII fragment of Drosophila hsp70 with nt -89 to +89 (relative to transcription start site) ligated to XhoI/HindIII Bluescript KS+ = pCII38. 248 bp SalI/XhoI fragment with 3' end of hsp70 ligated into SalI of Bluescript KS+ = pCIII1. (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.
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