Basic Vector Information
- Vector Name:
- pMpGWB431
- Antibiotic Resistance:
- Streptomycin
- Length:
- 12745 bp
- Type:
- Binary vector
- Replication origin:
- ori
- Source/Author:
- Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y, Shikanai T, Kohchi T.
- Promoter:
- CaMV35S(enhanced)
pMpGWB431 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 ℃.
pMpGWB431 vector Sequence
LOCUS 40924_31935 12745 bp DNA circular SYN 18-DEC-2018
DEFINITION Binary vector pMpGWB431 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12745)
AUTHORS Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y,
Shikanai T, Kohchi T.
TITLE Development of Gateway Binary Vector Series with Four Different
Selection Markers for the Liverwort Marchantia polymorpha
JOURNAL PLoS ONE 10 (9), E0138876 (2015)
PUBMED 26406247
REFERENCE 2 (bases 1 to 12745)
AUTHORS Nishihama R, Ishizaki K, Kohchi T.
TITLE Direct Submission
JOURNAL Submitted (01-JUN-2015) Contact:Ryuichi Nishihama Kyoto University,
Graduate School of Biostudies; Kitashirakawa-oiwake-cho, Sakyo-ku,
Kyoto, Kyoto 606-8502, Japan URL :http://www.lif.kyoto-u.ac.jp/e/
REFERENCE 3 (bases 1 to 12745)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12745)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2015"; volume: "10"; issue: "9"; pages: "E0138876"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-JUN-2015) Contact:Ryuichi Nishihama Kyoto University, Graduate
School of Biostudies"; volume: " Kitashirakawa-oiwake-cho, Sakyo-ku,
Kyoto, Kyoto 606-8502, Japan URL :http"; pages:
"//www.lif.kyoto-u.ac.jp/e"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12745
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(54..78)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 281..297
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 323..447
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 484..514
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 568..1224
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1569..1871
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1915..2039)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
RBS 3522..3530
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 3676..3795
/codon_start=1
/label=hPEST
/note="PEST degradation sequence from mouse ornithine
decarboxylase"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
terminator 3820..4072
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(4106..4122)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4130..4146
/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(4154..4184)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4199..4220)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
polyA_signal complement(4305..4479)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(4539..5327)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(5396..6072)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
misc_feature complement(6267..6291)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 6812..7600
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 7849..8437
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(8623..8763)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(9107..9301)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9370..10440)
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(10872..11498)
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
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