Last data update: 12 September 2023 11:24 CEST
Plasmid name: pCi-N-TagBFP-DEST (LMBP 9842)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9842.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene) Entacmaea quadricolor red fluorescent protein DNA (eqFP578, RFP); bright monomeric blue fluorescent variant (TagBFP), N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Escherichia coli lac operon promoter; mutant (lacUV5) Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Chloramphenicol (cam) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Copy number: | High copy number |
| Host range: | Escherichia coli; use a ccdB-resistant strain for propagation Mammalian cells |
| Parental clone: | pCI |
| Further information: | The plasmid was created by cloning the TagBFP coding sequence and the Gateway recombination cassette into the pCI vector. TagBFP is a monomeric blue fluorescent protein generated by site-specific and random mutagenesis of TagRFP. TagBFP possesses bright blue fluorescence with excitation/emission maxima at 402 and 457 nm, characterized by high photostability and extremely high pH-stability. This mammalian expression vector contains a chimeric intron downstream of the hCMV-IE promoter and enhancer which is composed of the 5´-donor site from the first intron of the human β-globin gene and the branch and 3´-acceptor site from the intron between the leader and the body of an immunoglobulin gene heavy chain variable region. The sequences of the donor and acceptor sites, along with the branchpoint site, have been changed to match the consensus sequences for splicing. Transfection studies have demonstrated that the presence of an intron flanking the cDNA insert frequently increases the level of gene expression. The intron is located 5´ to the cDNA insert in order to prevent utilization of possible cryptic 5´-donor splice sites within the cDNA sequence. This Gateway destination vector is compatible with the human orfeome library which is also available at BCCM/GeneCorner. The nucleotide sequence of the parental pCI plasmid corresponds with Genbank accession number U47119.2. Other name of the plasmid is pCi N-BFP2 DEST. |
| EMBL Accession number: | U47119.2, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 09/05/2016 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | The plasmid was deposited by Prof. Dr H. McMahon(1). It was constructed by Dr S. Lucken-Ardjomande Hasler(1). (1) Medical Research Council - Laboratory of Molecular Biology, Neurobiology Division, Cambridge University, Cambridge, United Kingdom |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC. |
| 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 + ampicillin (100 μg/ml) + chloramphenicol (34 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 ampicillin. |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pCi-N-TagBFP-DEST (LMBP 9842) is available at BCCM/GeneCorner. The plasmid was deposited by Prof. Dr H. McMahon. |
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.