Last data update: 04 December 2020 04:17 CET
Plasmid name: pLoxGentrc (LMBP 9151)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
|GeneCorner sequence:||p9151.gb (View with Genome Compiler)|
|Cloned DNA:||Escherichia coli lac repressor gene (lacI)|
|Promoter:||Escherichia coli hybrid tryptophan/lacUV5 promoter (trc)
Escherichia coli lac repressor promoter; mutant (lacIq)
Escherichia coli lac operon promoter
|Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)|
|Terminator:||Escherichia coli rrnB operon T1 terminator
Escherichia coli rrnB operon T2 terminator
|Selection marker:||Ampicillin (amp)
|Replicon:||Escherichia coli plasmid pMB1 origin
Bacteriophage M13 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was constructed based on pLoxGen4 by digesting it with HindIII and religating the backbone after blunting, eliminating this restriction site. A 2266 bp region containing lacIq, the trc promoter, the multicloning site and the T1 and T2 terminator sequences was PCR-amplified from pTrc99A and ligated as a BstZ117I/NotI fragment into the EcoRI (blunted)/NotI opened intermediary vector.
pLoxGentrc is meant for chromosomal gene integration and expression in E. coli.
The nucleotide sequence of the pLoxGentrc vector corresponds with the EMBL Nucleotide Sequence Database accession number JF934954.1.
|EMBL Accession number:||JF934954.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||05/05/2014|
Primers used to amplify the 2266 bp region of pTrc99A containing lacIq, trc promoter, MCS and T1 and T2 terminator: Ptrcup: 5' CTGATGCCGCATAGTTAAGCCA 3' Ptrcdown: 5' ATAAGAATGCGGCCGCGGGTTATTGTCTCATGAGCGG 3' NotI
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: HincII, PvuII and XhoI.
As compared to the compiled nucleotide sequence, one HincII site probably at position 2715 could not be experimentally confirmed.
|History of deposit:||This plasmid was deposited by Prof. Dr Guillermo Gosset Lagarda(1). It was constructed by A. Sabido(1).
(1) Departamento de Ingenieria Celular y Biocatalisis, Instituto de Biotecnologia, Universidad Nacional Autonoma de Mexico, Cuernavaca, Mexico
|Plasmid reference:||Sabido et al., Plasmid 69 (2013), 16-23 [PMID: 22884755] [DOI: 10.1016/j.plasmid.2012.04.005]
|Related plasmid reference:||Palmeros et al., Gene 247 (2000), 255-264 [PMID: 10773465]
|Restricted distribution:||- BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + gentamicin (10 μg/ml)*|
|Cultivation remark:||*: selection of transformants on gentamicin; subsequent cultivation of a single colony in liquid medium with ampicillin.|
|Other culture collection numbers:||-|
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.