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GeneCorner plasmid details

Last data update: 30 October 2020 04:11 CET

Plasmid name: pGBdest-ccdB (LMBP 12024)

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Price category: Cat. 2 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p12024.gb (View with Genome Compiler)
Sequence
analysis results
Genecorner:

Sanger: ef30589141.fasta

Sanger: ef30589133.fasta

Cloned DNA: GoldenBac destination module
B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Promoter: Escherichia coli plasmid R388 class 1 integron Pc promoter (PcS)
Escherichia coli class 1 integron integrase promoter (intI1)
Ribosome
binding site:
-
Terminator: -
Selection marker: Neomycin (neo; kanamycin (kan))
Gentamicin (Gm)
Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use a ccdB-resistant strain for propagation
Insect cells
Parental clone: pGBdest
Further information: The parental plasmid pGBdest is based on the pKL vector from the MultiBac system.
This plasmid is a destination vector for the assembly of expression cassettes for co-expression of up to 15 recombinant genes using a Bac-to-Bac baculoviral expression system (GoldenBac).
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 pGBdest_ccdB.
EMBL Accession number: -
Latest sequence update: 29/11/2019
Sequence detail:
Nucleotide sequence of the BsaI destination cassette:

ACCTTGAGACC...<- LacZ alpha --...<- ccdB --...<- CamR --...GGTCTCAGTAT
^^^^ BsaI                                                  BsaI   ****

^^^^: BsaI overhang 01
****: BsaI overhang 16
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI/BglII, EcoRV and HincII/HindIII.

The region of the GoldenBac cassette was sequenced at GeneCorner: from the 5' end of the gentamycin resistance gene to the 3' end of the chloramphenicol resistance gene and from 5' end of the chloramphenicol resistance gene to halfway the Tn903 neomycin resistance gene.
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 + kanamycin (50 µg/ml) + chloramphenicol (34 µg/ml)* + gentamycin (12,5 µg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with kanamycin and gentamycin.
Other culture collection numbers: -

Refer in your Materials and Methods:

pGBdest-ccdB (LMBP 12024) 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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