Last data update: 21 January 2021 04:16 CET
Plasmid name: pAc373 (LMBP 2304)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Promoter:||Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin promoter (PH)
Escherichia coli lac operon promoter
|Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)|
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli|
|Further information:||This plasmid was constructed by cloning the Autographa californica nuclear polyhedrosis virus (AcNPV) EcoRI-I fragment into the unique EcoRI expression site of the pUC8 multilinker region. All the sites of this multilinker, except HindIII at nucleotide position 399, have been removed.
The polyhedrin gene (APH) was inactivated by deletion of the region -9 to +175 (relative to the ATG start codon), where the unique BamHI restriction site has been inserted for expression of foreign genes. Around this BamHI expression site, there are nine nucleotides of unidentified origin (CGAGATCCG), between the deletion ending at -8 and the sequence comprising the BamHI site at position +171.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
There are no cleavage sites for BglII, PstI, SmaI, SacI and XbaI.
Some of the unique restriction sites mentioned are possibly not unique, because the nucleotide sequence is not completely known.
Experimentally verified restriction sites in the AcNPV nucleotide sequence are: HindIII, BstEII, PvuII, XhoI, SalI, EcoRV, BamHI, KpnI and EcoRI.
The physical stock of this plasmid contains a strain E2 virus fragment, but in absence of the nucleotide sequence information, the sequence of virus strain C6 was used for the theoretical construction.
|EMBL Accession number:||-|
|Latest sequence update:||04/02/1993|
Nucleotide sequence around the BamHI expression site: -16 -8 +176 | | | 5' AAAAAAACC CGAGATCCG CGGATC CTTTCCT 3' --------- BamHI | 9 nucleotides of unknown origin
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Dr M. Summers(1).
(1) Texas A&M university Systems, Texas, USA
|Plasmid reference:||Smith et al., Proc. Natl. Acad. Sci. U.S.A. 82 (1985), 8404-8408 [PMID: 3878519]
|Related plasmid reference:||Hooft Van Iddekinge et al., Virology 131 (1983), 561-565 [PMID: 18639177]
Possee et al., Virology 185 (1991), 229-241 [PMID: 1926775]
Smith et al., Mol. Cell. Biol. 3 (1983), 2156-2165 [PMID: 6318086]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pAc373 (LMBP 2304) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Summers and was published in Smith et al., 1985.|
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.