ID   PC4RO3     preliminary; circular DNA; SYN; 10800 BP.
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AC   ATCC77319;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Drosophila/E.coli plasmid vector pC4ro3 - incomplete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RC   pro2 from pBluescript KS+ & pro1
RC   pC4ro1 from pro2 & pC4cat
RC   pC4ro2 from pC4ro1
RC   pC4ro3 from pC4ro2 & pSL1180
RC   BstHsp from pBluescript KS+ & p56H8, Drosophila hsp70 gene promoter
RC   BstHspluc from BstHsp & pSV232AL-Adelta5', luc gene
RC   pC4ro3Hsp luc from pC4ro3 & BstHspluc
RA   Lockett T.J., Lewy D., Holmes P., Medveczky K., Saint R.;
RT   "The rough (ro+) gene as a dominant P-element marker in germ line
RT   transformation of Drosophila melanogaster";
RL   Gene 114:187-193(1992).
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RN   [2]
RC   pro1 from pUC18 & ro' mutation
RA   Saint R.;
RT   ;
RL   Unpublished (1992).
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CC   Vector containing a non-autonomous P-element and incorporating the
CC   rough gene (ro) as a dominant marker for P-element mediated gene
CC   transfer into the germ line of Drosophila melanogaster.
CC   The order of the major features in this plasmid is: HindIII -
CC   P-element/Carnegie 4/P-element - XhoI/MCS/SphI - ro gene.
CC   The SphI/HindIII fragment corresponding to the rough gene contains the
CC   following restriction sites (kb from the 5' end): BamHI--3.02;
CC   BglII--5.39; EcoRI--0.91, 4.91; SacI--0.57, 4.19, 5.86; SalI--1.80,
CC   3.91; SmaI--4.43. (personal communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   SalI--8.6, 2.2; HindIII--7.7, 3.1; PstI--10.8. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pC4ro3)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(Drosophila melanogaster)(E.coli)
CC   HO (E.coli TG1)(E.coli RR1)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. Carnegie 4 3630bp
FT                   -> pCar4
FT                   1. pCar4 3600bp
FT                   2. Tn9 TaqI-TaqI 773bp 215..988, cat gene/#V00622
FT                   -> pC4cat
FT                   1. pUC18 remove SphI-SalI 8bp 410..418, MCS/2678bp
FT                   2. D.melanogaster SphI-XhoI 6800bp, ro gene mutation
FT                   -> pro1 9700bp
FT                   1. pBluescript KS+ remove XbaI-HindIII 42bp 678..720,
FT                   \ MCS/2922bp
FT                   2. pro1 HindIII-XbaI 6800bp 400..410..-..418..424,
FT                   \ ro gene mutation
FT                   -> pro2 9700bp
FT                   1. pro2 KpnI 9700bp 443..443
FT                   T4 DNA polymerase
FT                   XbaI-XbaI 6800bp, ro gene mutation
FT                   2. pC4cat EcoRI-EcoRI
FT                   Klenow:Klenow
FT                   XbaI-XbaI, Carnegie 4 fragment
FT                   -> pC4ro1
FT                   1. pC4ro1 remove XbaI-PstI, MCS/10800bp
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pC4ro2 10800bp
FT                   1. pC4ro2 remove small XhoI-SphI, 10600bp
FT                   2. pSL1180 XhoI-SphI 154bp 2916..3070, MCS
FT                   -> pC4ro3 10800bp"
FT   misc_binding    0..0
FT                   /note="MCS XhoI-SplI-MluI-PstI-AvrII-StuI-HpaI-KpnI-
FT                   XbaI-SphI"
FT   misc_binding    0..0
FT                   /note="SIT unique XhoI-SplI-MluI-PstI-AvrII-StuI-
FT                   HpaI-KpnI-XbaI-SphI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
//