Last data update: 24 January 2024 16:39 CET
Plasmid name: pBlueBacmsGCB1His (LMBP 4467)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4467.gb
(View with Genome Compiler) p4467.txt p4467.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse guanylate cyclase 1 cDNA (mGC); soluble form, β1 subunit (msGCB1) = β3 (msGucy1b3) 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 msGCB1 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 β1 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 LT727215.1. The nucleotide sequence of the msGCB1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF020339.1. Other name of the plasmid is pBlueBac4.5/V5-msGCBeta1-His. |
| EMBL Accession number: | AF020339.1, view at EMBL, GenBank, DDBJ LT727215.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/01/2002 |
| Sequence detail: | Sequence at the 3' end of the msGCB1 insert:
msGCB1 620| |<- V5 epitope
5' GAG.GAT.GAA.AAC.AAG.GGC.AAT.TCG.AAG.CCT.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: BglII, HindIII and XmnI. |
| 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: | Instruction Manual (Invitrogen) |
| 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: |
pBlueBacmsGCB1His (LMBP 4467) 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.