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ID   PHSREM3    preliminary; circular DNA; SYN; 3400 BP.
AC   ATCC37644;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   E. coli plasmid vector pHSREM3 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pHSREM1 from pBluescript KS+ & Drosophila hsp70 gene prom/term
RC   pHSREM2 from pBluescript KS- & Drosophila hsp70 gene prom/term
RC   pHSREM3 from pBluescript SK+ & Drosophila hsp70 gene prom/term
RC   pHSREM4 from pBluescript SK- & Drosophila hsp70 gene prom/term
RA   Knipple D.C., Marsella-Herrick P.;
RT   "Versatile plasmid vectors for the construction, analysis and
RT   heat-inducible expression of hybrid genes in eukaryotic cells";
RL   Nucleic Acids Res. 16:7748-7748(1988).
CC   A KpnI/SacI double digest releases the hsp70 transcription module, for
CC   subsequent ligation into other plasmids such as ones containing
CC   Drosophila P elements. KpnI BglII NotI XhoI - promoter - MCS -
CC   terminator - SpeI XbaI NotI EagI BstXI SacII SacI.
CC   One of a series (ATCC 37642-37645) of plasmid vectors for expressing
CC   genes under the control of the Drosophila hsp70 promoter & terminator.
CC   The transcription module is flanked by restriction sites (facilitating
CC   its removal), T7 and T3 promoters (for in vitro RNA synthesis), &
CC   primers for Sanger sequencing.  There is also an M13 origin of
CC   replication.
CC   Restriction digests of the clone give the following sizes (kb):
CC   PstI--3.5; HindIII--3.4; BamHI--3.4. (ATCC staff)
CC   The KpnI/SacI fragment of pHSREM1 (ATCC 37642) containing the hsp70
CC   transcription module was ligated into KpnI/SacI digested Bluescript
CC   SK+. (personal communication)
CC   Major Drosophila hsp70 regulator elements are: 2 heat shock consensus
CC   elements (-85 to -72 & -62 to -49),a TATA box (-33 to -26) &
CC   transcription initiation site (+1) in the 5' segment; the termination
CC   triplet and polyA site in the 3' segment. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli HB101)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pBluescript SK+ remove PstI-XhoI 35bp
FT                   \ 706..741, MCS/2929bp
FT                   2. Drosophila XhoI-PstI, hsp70 gene promoter
FT                   -> plasmid
FT                   1. plasmid remove SalI-SpeI 49bp 735..684, MCS/ bp
FT                   2. Drosophila SalI-SpeI, hsp70 gene terminator
FT                   -> pHSREM3 3400bp"
FT   misc_binding    0..0
FT                   /note="MCS SalI-ClaI-HindIII-EcoRV-EcoRI-PstI-SmaI-
FT                   BamHI-SmaI-PstI-EcoRI-EcoRV-HindIII-PstI"
FT   misc_binding    0..0
FT                   /note="SIT unique PstI-HindIII-EcoRV-EcoRI-SmaI-
FT                   BamHI-ClaI-SalI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage M13"
FT   promoter        0..0
FT                   /note="PRO Drosophila hsp70 gene"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T3"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T7"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;