GeneCorner plasmid details

Last data update: 28 October 2020 04:17 CET

Plasmid name: pGB-04-05-ccdB (LMBP 12028)

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Price category: Cat. 2 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: (View with Genome Compiler)
analysis results

Sanger: ef30589139.fasta

Sanger: ef30589146.fasta

Cloned DNA: GoldenBac entry module
B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Strep-tag III (Twin-Strep-tag)
Human rhinovirus 3C protease cleavage site (PreScission site, PRS)
Histidine tag (His-tag)
Promoter: Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin promoter (PH)
Escherichia coli class 1 integron integrase promoter (intI1)
Escherichia coli plasmid R388 class 1 integron Pc promoter (PcS)
binding site:
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Gentamicin (Gm)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use a ccdB-resistant strain for propagation
Insect cells
Parental clone: pGB
Further information: The plasmid was constructed by cloning a cassette consisting of BsaI-pAcNPV-PH-Strep-Prescission-ccdB-His-SV40-BsaI into the pGB vector. This pGB vector is based on pACEBac1 from the MultiBac system.
This plasmid is an entry cloning vector for single gene expression of recombinant proteins in insect cells using a Bac-to-Bac baculoviral expression system (GoldenBac). With 16 different overhangs, the system allows for co-expression of up to 15 genes after assembly into a destination vector.
GoldenBac is a modular cloning system designed for insect cells which makes use of a single type IIS restriction endonuclease (BsaI).
The BsaI cassette is flanked by Tn7 transposon sequences.
Other name of the plasmid is pGB-04;05_ccdB.
EMBL Accession number: -
Latest sequence update: 29/11/2019
Sequence detail:
Nucleotide sequence of the BsaI cassette:

GGTCTCATCAG...-- pPH ->....-- SV40 polyA ->...CTGCTGAGACC
BsaI   ^^^^                                   **** BsaI

^^^^: BsaI overhang 04
****: BsaI overhang 05
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AgeI/BglII, NcoI and NotI/SpeI.

The region of the GoldenBac cassette was sequenced at GeneCorner: from the 5' end of the gentamycin resistance gene to the Tn7 recombination sequence.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr D. Soroldoni(1). It was constructed by Dr J. Neuhold(1).
(1) Vienna Biocenter Core Facilities GmbH, Vienna, Austria
Plasmid reference: Neuhold et al., BMC Biotechnology 20 (2020), 26 [PMID: 32398045] [DOI: 10.1186/s12896-020-00616-z]
Related plasmid reference: Lampropoulos et al., PLoS ONE 8 (2013), e83043 [PMID: 24376629] [DOI: 10.1371/journal.pone.0083043]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12xB DB3.1
Host reference: -
Related host reference: Bernard et al., J. Mol. Biol. 226 (1992), 735-745 [PMID: 1324324]
Cultivation medium: LB-Lennox + gentamicin (12,5 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pGB-04-05-ccdB (LMBP 12028) is available at BCCM/GeneCorner. This plasmid was deposited by Dr D. Soroldoni and was published in Neuhold et al., 2020.

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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