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GeneCorner plasmid details

Last data update: 30 November 2020 04:30 CET

Plasmid name: pBlueBacmsGCA1His (LMBP 4470)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4470.gb (View with Genome Compiler)
p4470.txt
p4470.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse guanylate cyclase 1 cDNA (mGC); soluble form, α1 subunit (msGCA1) = α3 (msGucy1a3)
V5 epitope; C-terminal
Histidine tag (His-tag); C-terminal
Escherichia coli lac Z gene (lacZ); 5' fragment
Autographa californica nuclear polyhedrosis virus (AcNPV) ORF 1629; 3' fragment
Promoter: Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin promoter (PH)
Autographa californica nuclear polyhedrosis virus (AcNPV) EcoRI-T large promoter (ETL)
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Insect cells; e.g. Sf9 cells
Parental clone: pBlueBac4.5/V5-His-TOPO
Further information: The plasmid was constructed by inserting a TaqI amplified PCR fragment, containing the msGCA1 coding sequence, in the topoisomerase I-activated pBlueBac4.5/V5-His-TOPO vector, in phase with the C-terminal His-tag and V5 epitope.
After cotransfection with Bac-N-Blue‚ Linear DNA, the plasmid can be used for mouse soluble guanylate cyclase α1 expression in insect cells. The 5' lacZ fragment recombines to the lacZ sequence in the Bac-N-Blue DNA and produces blue recombinant plaques for easy selection. The 3' ORF 1629 fragment recombines and restores the essential ORF 1629 for production of viable, recombinant virus.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727217.1.
The nucleotide sequence of the msGCA1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF297082.1.
Other name of the plasmid is pBlueBac4.5/V5-msGCAlpha1-His.
EMBL Accession number: AF297082.1, view at EMBL, GenBank, DDBJ
LT727217.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 10/01/2002
Sequence detail:
Sequence at the 3' end of the msGCA1 insert:

       msGCA1  671|                               |<-  V5 epitope
5' TCA.GGG.GTA.GAT.AAG.GGC.AAT.TCG.AAG.CTT.AGG.CCT.GGT.AAG.CCT.ATC.CCT.AAC.CCT.
                                   HindIII

                         ->|           |<-   His-tag         ->|   
CTC.CTC.GGT.CTC.GAT.TCT.ACG.CGT.ACC.GGT.CAT.CAT.CAC.CAT.CAC.CAT.TGA.CGTCTCTAGCTTGGAGTCGACAA 3'
                                AgeI                                               SalI
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BstXI, EcoRV and SacI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr P. Brouckaert(1) (2). It was constructed by M. Buys(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 TOP10
Host reference: Invitrogen Instruction Manual
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pBlueBacmsGCA1His (LMBP 4470) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Brouckaert .

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.

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