Basic Vector Information
- Vector Name:
- pMF270
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8122 bp
- Type:
- Neurospora expression vector
- Replication origin:
- ori
- Source/Author:
- Honda S, Selker EU.
pMF270 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pMF270 vector Sequence
LOCUS 40924_30710 8122 bp DNA circular SYN 18-DEC-2018 DEFINITION Neurospora expression vector pMF270, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8122) AUTHORS Honda S, Selker EU. TITLE Tools for fungal proteomics: multifunctional neurospora vectors for gene replacement, protein expression and protein purification JOURNAL Genetics 182 (1), 11-23 (2009) PUBMED 19171944 REFERENCE 2 (bases 1 to 8122) AUTHORS Honda S, Selker EU. TITLE Direct Submission JOURNAL Submitted (11-NOV-2008) Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA REFERENCE 3 (bases 1 to 8122) TITLE Direct Submission REFERENCE 4 (bases 1 to 8122) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genetics"; date: "2009"; volume: "182"; issue: "1"; pages: "11-23" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (11-NOV-2008) Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..8122 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 363..467 /label=AmpR promoter CDS 468..1325 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1499..2087 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 2345..2363 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2638..2649 /codon_start=1 /label=Factor Xa site /note="Factor Xa recognition and cleavage site" /translation="IEGR" primer_bind 4981..4997 /label=SK primer /note="common sequencing primer, one of multiple similar variants" CDS 5019..5108 /codon_start=1 /label=3xHA /note="three tandem HA epitope tags" /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA" terminator 5141..5328 /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(5396..5412) /label=KS primer /note="common sequencing primer, one of multiple similar variants" promoter complement(5442..5460) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter complement(join(8121..8122,1..17)) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase"
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