Last data update: 24 January 2024 16:39 CET
Plasmid name: pGBdest-ccdB (LMBP 12024)
New search | Print data sheet |
Price category: | Cat. 3 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p12024.gb
(View with Genome Compiler) p12024.txt p12024.pdf |
Sequence analysis results Genecorner: |
Sanger: ef30589141.fasta Sanger: ef30589133.fasta NGS: a03-gc-may2021.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. This plasmid has also been fully sequenced. |
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 Biotechnol. 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.