Last data update: 24 January 2024 16:39 CET
Plasmid name: pBlueBachsGCA1His (LMBP 4464)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p4464.gb
(View with Genome Compiler) p4464.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Human guanylate cyclase 1 soluble subunit alpha 1 cDNA (GUCY1A1, GC-SA3, GUC1A3, GUCY1A3, sGCA1, GeneID 2982) 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 human soluble guanylate cyclase α1 coding sequence (hsGCA1), in phase with the C-terminal His-tag and V5 epitope in the topoisomerase I-activated pBlueBac4.5/V5-His-TOPO vector. After cotransfection with Bac-N-Blue‚ Linear DNA the plasmid can be used for hsGCA1 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' ORF1629 fragment recombines and restores the essential ORF1629 for production of viable, recombinant virus. |
EMBL Accession number: | - |
Latest sequence update: | 20/12/2001 |
Sequence detail: | Sequence at the 3' end of the inserted gene: hsGCA1 690| |<- V5 epitope 5' GGA.ATA.GAT.AAG.GGC.AAT.TCG.AAG.CTT.AGG.CCT.GGT.AAG.CCT.ATC.CCT.AAC.CCT.CTC.CTC.GGT.CTC. HindIII ->| |<- His-tag ->| 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: AvaI, HindIII and NcoI/XbaI. |
Class: | Recombinant plasmid |
Type: | Plasmid |
History of deposit: | This plasmid was deposited by Prof. Dr P. Brouckaert(1)(2) and 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: |
pBlueBachsGCA1His (LMBP 4464) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Brouckaert and constructed by M. Buys. |
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.