Basic Vector Information
p3xHA NPTII 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 ℃.
p3xHA NPTII vector Sequence
LOCUS 40924_2732 5405 bp DNA circular SYN 18-DEC-2018
DEFINITION Moss transformation vector p3xHA NPTII DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5405)
AUTHORS Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
TITLE Moss transformation vectors
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5405)
AUTHORS Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
TITLE Direct Submission
JOURNAL Submitted (26-NOV-2015) Contact:Yosuke Tamada National Institute for
Basic Biology, Division of Evolutionary Biology; 38 Nishigonaka,
Myodaiji, Okazaki, Aichi 444-8585, Japan URL
:http://www.nibb.ac.jp/evodevo/
REFERENCE 3 (bases 1 to 5405)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5405)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-NOV-2015) Contact:Yosuke Tamada National Institute for Basic
Biology, Division of Evolutionary Biology; 38 Nishigonaka, Myodaiji,
Okazaki, Aichi 444-8585, Japan URL :http://www.nibb.ac.jp/evodevo/"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5405
/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 600..616
/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 670..686
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 701..790
/codon_start=1
/label=3xHA
/note="three tandem HA epitope tags"
/translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
misc_recomb 822..855
/label=loxP
/note="loxP"
protein_bind 822..855
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
terminator 860..1111
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 1194..1541
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 1562..2353
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 2426..2602
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
protein_bind complement(3091..3124)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter complement(3217..3235)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3256..3272)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3280..3296)
/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(3304..3334)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3349..3370)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3658..4246)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4420..5277)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5278..5382)
/label=AmpR promoter
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