Basic Vector Information
- Vector Name:
- pNW55
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3224 bp
- Type:
- Plant Expression, RNAi ; Plant gene inactivation
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7
- 3' Primer:
- T3
pNW55 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 ℃.
pNW55 vector Sequence
LOCUS 40924_33677 3224 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3224)
AUTHORS Warthmann N, Chen H, Ossowski S, Weigel D, Herve P
TITLE Highly specific gene silencing by artificial miRNAs in rice.
JOURNAL PLoS One. 2008 . 3(3):e1829.
PUBMED 18350165
REFERENCE 2 (bases 1 to 3224)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 3224)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.
2008 . 3(3):e1829."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3224
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 603..619
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 677..693
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(988..1004)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(1033..1051)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1072..1088)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1096..1112)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(1120..1150)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1165..1186)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1303..1320)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1474..2062)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2236..3093)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3094..3198)
/label=AmpR promoter
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