Basic Vector Information
- Vector Name:
- pMN20
- Antibiotic Resistance:
- Spectinomycin
- Length:
- 11073 bp
- Type:
- Plant Expression
- Replication origin:
- ori
- Selection Marker:
- kanamycin
- Copy Number:
- High Copy
- Promoter:
- CaMV35S(enhanced)
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- M13 reverse
pMN20 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 ℃.
pMN20 vector Sequence
LOCUS 40924_31165 11073 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 11073)
AUTHORS Weigel D, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK,
Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM,
Nguyen JT, Sato S, Wang ZY, Xia Y, Dixon RA, Harrison MJ, Lamb CJ,
Yanofsky MF, Chory J
TITLE Activation tagging in Arabidopsis.
JOURNAL Plant Physiol. 2000 Apr . 122(4):1003-13.
PUBMED 10759496
REFERENCE 2 (bases 1 to 11073)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11073)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Physiol. 2000 Apr . 122(4):1003-13."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11073
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(60..76)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2053..2069)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2053..2069)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(2109..2125)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2109..2125)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2122..2144)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2133..2149
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(2157..2187)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2202..2223)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 2388..2412
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 3712..4338
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 4775..5839
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 5908..6102
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(6338..6360)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 6446..6586
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(6601..6618)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6772..7360)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7607..8395)
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 8923..8947
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(9045..9219)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9276..10064)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(10130..10808)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 11037..11053
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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