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ID   PRSVPAP    preliminary; circular DNA; SYN; 10100 BP.
AC   ATCC77130;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   Vertebrate/E.coli phagemid vector pRSVPAP - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pRSVADH from pWX008 & pRc/RSV
RC   pRSVPAP from pSV2Apap & pRc/RSV
RC   plasmid from pM-MuLV-SV-lacZ & pSK+
RC   plasmid from plasmid & pECE
RC   pRSVlacZ from plasmid & pRc/RSV
RC   pCMVlacZ from pM-MuLV-SV-lacZ
RC   pCMVADH from pWX008
RA   Lin W.C., Culp L.A.;
RT   "Selectable plasmid vectors with alternative and ultrasensitive
RT   histochemical marker genes";
RL   Biotechniques 11:344-351(1991).
CC   Transient expression experiments should be performed to examine the
CC   RSV promoter efficiency in different cell lines.   The RSV promoter
CC   can be replaced by excising with a BglII/HindIII digest. (personal
CC   communication)
CC   Shuttle reporter plasmid permitting visual detection of activity by
CC   histochemical staining. [1]
CC   The order of the major features in pRc/RSV is:RSV LTR - MCS - bovine
CC   growth hormone polyadenylation signal - M13 ori - SV40 early promoter
CC   - neoR - SV40 polyadenylation signal - pMB1 ori - bla. [1]
CC   pRSVlacZII (ATCC 77129), pRSVPAP (ATCC 77130) and pRSVADH (ATCC 77131)
CC   provide distinct color-staining reactions (aqua blue, red, and
CC   blue-black respectively) to permit simultaneous analysis of multiple
CC   lineages. [1]
CC   Presence of alkaline phosphatase activity is used to follow cell
CC   lineage in culture or in situ. [1]
CC   Constructed by cloning the HindIII/XbaI fragment from pSV2Apap into
CC   pRc/RSV. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--6.1, 4.0; HindIII--10.1; BglII--10.1; KpnI--5.0, 2.8, 2.3;
CC   XbaI--10.1. (ATCC staff)
CC   A (GT)n repeat. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   HO (E.coli DH5alpha)(vertebrate cells)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pRcRSV from Invitrogen)(pSV2Apap)
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pSV2Apap HindIII-XbaI 5000bp, ALP gene
FT                   2. pRc/RSV large XbaI-HindIII
FT                   -> pRSVPAP 10100bp"
FT   CDS             0..0
FT                   /note="GEN human placenta alkaline phosphatase ALP
FT                   gene, from pSV2Apap"
FT   promoter        0..0
FT                   /note="PRO RSV LTR, from pRcRSV"
FT   polyA_signal    0..0
FT                   /note="PLA E. coli BGH gene, from pRcRSV"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage M13, from pRcRSV"
FT   promoter        0..0
FT                   /note="PRO SV40 early gene, from pRcRSV"
FT   CDS             0..0
FT                   /note="ANT E. coli neomycin phosphotransferase gene
FT                   (NPT); neomycin resistance gene (neo)
FT                   mammal G418 resistance gene (G418), from pRcRSV"
FT   polyA_signal    0..0
FT                   /note="PLA SV40 early gene, from pRcRSV"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322),
FT                   from pRcRSV"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp), from pRcRSV"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;