GREAT AT SMALL THINGS

0

GeneCorner plasmid details

Last data update: 24 October 2020 04:12 CEST

Plasmid name: pBlueBachsGCA1His (LMBP 4464)

Add to cart

New search

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: 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:

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.

New search