Basic Vector Information
- Vector Name:
- pN-3xHA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6969 bp
- Type:
- Neurospora expression vector
- Replication origin:
- ori
- Source/Author:
- Honda S, Selker EU.
pN-3xHA 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 ℃.
pN-3xHA vector Sequence
LOCUS 40924_32953 6969 bp DNA circular SYN 18-DEC-2018
DEFINITION Neurospora expression vector pN-3xHA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6969)
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 6969)
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 6969)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6969)
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..6969
/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"
promoter 4008..4026
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 4056..4072
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 4118..4207
/codon_start=1
/label=3xHA
/note="three tandem HA epitope tags"
/translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
promoter complement(4289..4307)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(join(6968..6969,1..17))
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
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