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ID   PC4RO3     preliminary; circular DNA; SYN; 10800 BP.
AC   ATCC77319;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Drosophila/E.coli plasmid vector pC4ro3 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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).
RN   [2]
RC   pro1 from pUC18 & ro' mutation
RA   Saint R.;
RT   ;
RL   Unpublished (1992).
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   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 ()
FH   Key             Location/Qualifiers
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)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;