Last data update: 31 October 2020 04:21 CET
Plasmid name: BH20 (LMBP 7765)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Phage MS2 assembly protein gene (A)
Phage MS2 coat protein gene (COAT)
RT-PCR positive control DNA
Histidine tag (His-tag); N-terminal
Thrombin recognition site; N-terminal
Histidine tag (His-tag); C-terminal
Escherichia coli lac repressor gene (lacI)
|Promoter:||Escherichia coli lac repressor promoter (lacI)
Phage T7 gene 10 promoter (T7g10) with lac operator (T7lac)
|Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)|
|Terminator:||Phage T7 gene 10 terminator (T7g10)|
|Selection marker:||Neomycin (neo; kanamycin (kan))|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was created by ligating a 178 bp DNA sequence, specific for positive control of RT-PCR, into the HindIII site of the AU2 plasmid.
The plasmid is useful for the production of armored RNA.
The His-tags, the thrombin recognition site and the S-tag are not in frame with the phage MS2 A and COAT genes.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727566.1.
The nucleotide sequence of the pET-30b(+) vector part corresponds with the pET-30b(+) sequence file from Novagen (version 25/5/2002).
The nucleotide sequences upstream of the f1 origin and downstream from the T7 promoter were sequenced by GeneCorner.
The nucleotide sequence of the pMS27 insert corresponds with the BCCM/GeneCorner sequence analysis results and with the sequence analysis results from the depositor. However, the 'C' nucleotide at position 291 of the COAT protein cds was originally a 'T' in the sequence file of pMS27 (LMBP 5349). This nucleotide change concerns a silent mutation.
|EMBL Accession number:||LT727566.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||10/09/2012|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BglI, HincII, HindIII/KpnI, PvuII and XbaI.|
|History of deposit:||This plasmid was deposited by Dr V. Jestin(1).
(1) VIPAC unit, National reference laboratory on avian influenza & Newcastle disease, Ploufragan, France
|Related plasmid reference:||Yu et al., J. Clin. Microbiol. 46 (2008), 837-841 [PMID: 18160451]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli B BL21(DE3)|
|Host reference:||Studier and Moffatt, J. Mol. Biol. 189 (1986), 113-130 [PMID: 3537305] [DOI: 10.1016/0022-2836(86)90385-2]
|Related host reference:||Studier et al., J. Mol. Biol. 394 (2009), 653-680 [PMID: 19765592] [DOI: 10.1016/j.jmb.2009.09.021]
Jeong et al., J. Mol. Biol. 394 (2009), 644-652 [PMID: 19786035] [DOI: 10.1016/j.jmb.2009.09.052]
Daegelen et al., J. Mol. Biol. 394 (2009), 634-643 [PMID: 19765591] [DOI: 10.1016/j.jmb.2009.09.022]
|Cultivation medium:||LB-Lennox + kanamycin (50 μg/ml)|
|Cultivation remark:||Take a low-copy-plasmid yield into account while choosing a plasmid DNA isolation procedure.|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|BH20 (LMBP 7765) is available at BCCM/GeneCorner. This plasmid was deposited by Dr V. Jestin.|
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.