pBC12BI [4053B] (ATCC® 67617)

Applications: contains sequence preproinsulin 2expression vectorshuttle vectorvector permitting construction of fusion proteins  /  Depositors: Hoffmann-La Roche Ltd., PC Watkins, Hoffmann-La Roche Ltd.

Designations pBC12BI [4053B]
Permits and Restrictions

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Depositors Hoffmann-La Roche Ltd., PC Watkins, Hoffmann-La Roche Ltd.
U.S. Patent
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
Biosafety Level 1
Vector Information
Size (kb): 4.0700001716613770
Vector: pBC12BI (plasmid)
Promoters: Promoter RSV LTR
Construction: pXF3, pBC81delta4, SV40
Construct size (kb): 4.099999904632568
Features: marker(s): ampR
promoter: RSV LTR
replicon: pMB1, SV40
terminator: rat insulin II
enhancer: RSV LTR
contains sequence preproinsulin 2
expression vector
shuttle vector
vector permitting construction of fusion proteins
Restriction digests of the clone give the following sizes (kb): BamHI--4.1; EcoRI--2.8, 0.78, 0.44; HindIII--4.1; BstEII--4.1.
This was constructed from pXF3 (pBR322 minus the poison sequence), a SV40 ori, and a rat preproinsulin genomic sequence under the control of RSV LTR (pBC81delta4 cut at BgIII and BalI and inserted at the BamHI and ClaI sites).
cDNA sequences with a translation initiation codon are usually inserted between the unique HindIII and BamHI sites (5' to 3'). Polyadenylation signals (and possibly carboxy terminal amino acids) are contributed by the preproinsulin gene.
Genomic sequences are normally inserted between unique HindIII and SmaI sites (5' to 3'). This deletes the preproinsulin gene from the vector.
cDNA clones without an initiation codon may be cloned in frame into the unique BamHI site 3' to the AUG of the rat preproinsulin gene. This results in 5 - 6 additional amino terminal amino acids from the insulin signal peptide.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C

Cullen BR. Use of eukaryotic expression technology in the functional analysis of cloned genes. Methods Enzymol. 152: 684-704, 1987. PubMed: 3657593

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