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ID PBC12BI preliminary; circular DNA; SYN; 4070 BP.
DT 01-JUL-1993 (Rel. 7, Created)
DT 01-APR-1995 (Rel. 11, Last updated, Version 1)
DE Vertebrate/E.coli plasmid vector pBC12BI - incomplete.
KW cloning vector.
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
RC pGJ20 from pGJ15 & pBR-F16, FSV LTR
RC pBC20It from pBR322 & pGJ20, FSV LTR
RC pBC20Is from pBC20It
RC pBC21It from pBC20It
RC pBC21Is from pBC20Is
RC pBC12BI from pBC12MI & RSV LTR
RC pKR2/LTR-R from pPB12 & SR-B LTR
RC pKR2/RAV-0 from pPB12 & RAV-0 LTR
RC pKR2/FSV from pPB12 & FSV LTR
RA Cullen B.R., Raymond K., Ju G.;
RT "Transcriptional activity of avian retroviral long terminal
RT repeats directly correlates with enhancer activity";
RL J. Virol. 53:515-521(1985).
RC from pBC12BI
RC from p91023(B)
RA Cullen B.R.;
RT "Use of eukaryotic expression technology in the functional
RT analysis of cloned genes";
RL Meth. Enzymol. 152:684-704(1987).
RC pYM141 from pIN-II series & E.coli lpp gene
RA Morinaga Y., Franceschini T., Inouye S., Inouye M.;
RT "Improvement of oligonucleotide-directed site-specific mutagenesis
RT using double-stranded plasmid DNA";
RL Biotechnology 2:636-639(1984).
RC p91023(B) from pQ2 & mouse DHFR gene
RA Kaufman R.J.;
RT "Identification of the components necessary for adenovirus
RT translational control and their utilization in cDNA expression
RL Proc. Natl. Acad. Sci. U.S.A. 82:689-693(1985).
CC cDNA clones without an initiation codon may be cloned in frame into
CC the unique BamHI site 3' to the AUG of the rat preproinsulin gene.
CC This results in 5 - 6 additional amino terminal amino acids from the
CC insulin signal peptide. 
CC Genomic sequences are normally inserted between unique HindIII and
CC SmaI sites (5' to 3'). This deletes the preproinsulin gene from the
CC vector. 
CC cDNA sequences with a translation initiation codon are usually
CC inserted between the unique HindIII and BamHI sites (5' to 3'). 
CC Polyadenylation signals (and possibly carboxy terminal amino acids)
CC are contributed by the preproinsulin gene. 
CC This was constructed from pXF3 (pBR322 minus the poison sequence), a
CC SV40 ori, and a rat preproinsulin genomic sequence under the control
CC of RSV LTR (pBC81delta4 cut at BglII and BalI and inserted at the
CC BamHI and ClaI sites). 
CC Restriction digests of the clone give the following sizes (kb):
CC BamHI--4.1; EcoRI--2.8,0.78,0.44; HindIII--4.1; BstEII--4.1. (ATCC
CC Medium is 1227 LB plus ampicillin.
CC NM (pBC12BI)
CC CM (no)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (ATCC)
CC HO (E.coli RR1)(vertebrate cells)(E.coli)
CC HO (monkey COS-1 cell line)(mammal cell line)(avian cell line)
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. lambda
FT -> lambda SRB-td8
FT 1. ALV SR-B
FT -> pLD6
FT 1. pBR322
FT 2. lambda SRB-td8
FT 3. pLD6, ALV SR-B
FT -> pBC81
FT 1. pBC81
FT -> pBC81delta3
FT 1. pBC81delta3
FT -> pBC81delta4
FT 1. pXf3 remove small NruI-SphI
FT 2. SV40 HindIII-SphI 200bp, SV40 origin
FT -> pXF3/ori
FT 1. pBC81delta4 BglII-BalI, SV40/RSV LTR
FT 2. pXF3/ori remove small BamHI-ClaI
FT :blunt end
FT -> pBC12BI"
FT misc_binding 0..0
FT /note="MCS BamHI-HindIII-SmaI-HindIII-BamHI"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT rep_origin 0..0
FT /note="ORI SV40"
FT promoter 0..0
FT /note="PRO RSV LTR"
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;