Basic Vector Information
- Vector Name:
- pMW399
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 12509 bp
- Type:
- Plant transient expression vector
- Replication origin:
- oriV
- Host:
- Plants
- Source/Author:
- Kagale S, Uzuhashi S, Wigness M, Bender T, Yang W, Borhan MH, Rozwadowski K.
- Promoter:
- CaMV 35S (enhanced)
pMW399 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 ℃.
pMW399 vector Sequence
LOCUS V004384 12509 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004384
VERSION V004384
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12509)
AUTHORS Kagale S, Uzuhashi S, Wigness M, Bender T, Yang W, Borhan MH,
Rozwadowski K.
TITLE TMV-Gate vectors: Gateway compatible tobacco mosaic virus based
expression vectors for functional analysis of proteins
JOURNAL Sci Rep 2, 874 (2012)
PUBMED 23166857
REFERENCE 2 (bases 1 to 12509)
AUTHORS Rozwadowski K, Kagale S.
TITLE Direct Submission
JOURNAL Submitted (15-OCT-2012) Molecular Genetics, Agriculture and
Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan S7N
0X2, Canada
REFERENCE 3 (bases 1 to 12509)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12509)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep 2,
874 (2012)"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-OCT-2012) Molecular Genetics, Agriculture and Agri-Food Canada,
107 Science Place, Saskatoon, Saskatchewan S7N 0X2, Canada"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12509
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 5..29
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
primer_bind 110..126
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter 230..899
/label="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
misc_feature 911..966
/label="TMV Omega"
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
CDS 968..4318
/codon_start=1
/gene="Rep"
/product="replicase"
/label="Rep"
/protein_id="AFX83526.1"
/translation="MAYTQTATTSALLDTVRGNNSLVNDLAKRRLYDTAVEEFNARDRR
PKVNFSKVISEEQTLIATRAYPEFQITFYNTQNAVHSLAGGLRSLELEYLMMQIPYGSL
TYDIGGNFASHLFKGRAYVHCCMPNLDVRDIMRHEGQKDSIELYLSRLERGGKTVPNFQ
KEAFDRYAEIPEDAVCHNTFQTCEHQPMQQSGRVYAIALHSIYDIPADEFGAALLRKNV
HTCYAAFHFSENLLLEDSCVNLDEINACFSRDGDKLTFSFASESTLNYCHSYSNILKYV
CKTYFPASNREVYMKEFLVTRVNTWFCKFSRIDTFLLYKGVAHKSVDSEQFYTAMEDAW
HYKKTLAMCNSERILLEDSSSVNYWFPKMRDMVIVPLFDISLETSKRTRKEVLVSKDFV
FTVLNHIRTYQAKALTYANVLSFVESIRSRVIINGVTARSEWDVDKSLLQSLSMTFYLH
TKLAVLKDDLLISKFSLGSKTVCQHVWDEISLAFGNAFPSVKERLLNRKLIRVAGDALE
IRVPDLYVTFHDRLVTEYKASVDMPALDIRKKMEETEVMYNALSELSVLRESDKFDVDV
FSQMCQSLEVDPMTAAKVIVAVMSNESGLTLTFERPTEANVALALQDQEKASEGALVVT
SREVEEPSMKGSMARGELQLAGLAGDHPESSYSKNEEIESLEQFHMATADSLIRKQMSS
IVYTGPIKVQQMKNFIDSLVASLSAAVSNLVKILKDTAAIDLETRQKFGVLDVASRKWL
IKPTAKSHAWGVVETHARKYHVALLEYDEQGVVTCDDWRRVAVSSESVVYSDMAKLRTL
RRLLRNGEPHVSSAKVVLVDGVPGCGKTKEILSRVNFDEDLILVPGKQAAEMIRRRANS
SGIIVATKDNVKTVDSFMMNFGKSTRCQFKRLFIDEGLMLHTGCVNFLVAMSLCEIAYV
YGDTQQIPYINRVSGFPYPAHFAKLEVDEVETRRTTLRCPADVTHYLNRRYEGFVMSTS
SVKKSVSQEMVGGAAVINPISKPLHGKILTFTQSDKEALLSRGYSDVHTVHEVQGETYS
DVSLVRLTPTPVSIIAGDSPHVLVALSRHTCSLKYYTVVMDPLVSIIRDLEKLSSYLLD
MYKVDAGTQ"
gene 968..4318
/gene="Rep"
/label="Rep"
CDS 4394..5812
/codon_start=1
/gene="RNA pol"
/product="RNA polymerase"
/label="RNA pol"
/protein_id="AFX83527.1"
/translation="MQFYYDKCLPGNSTMMNNFDAVTMRLTDISLNVKDCILDMSKSVA
APKDQIKPLIPMVRTAAEMPRQTGLLENLVAMIKRNFNAPELSGIIDIENTASLVVDKF
FDSYLLKEKRKPNKNVSLFSRESLNRWLEKQEQVTIGQLADFDFVDLPAVDQYRHMIKA
QPKQKLDTSIQTEYPALQTIVYHQKRSMQYRPLLVSLPGNYWTVLIRADFVFTRKTPAQ
IEDFFGDLDSHVPMDVLELDISKYDKSQNEFHCAVEYEIWRRLGFEDFLGEVWKQGHRK
TTLKDYTAGIKTCIWYQRKSGDVTTFIGNTVIIAACLASMLPMEKIIKGAFCGDDSLLY
FPKGCEFPDVQHSANLMWNFEAKLFKKQYGYFCGRYVIHHDRGCIVYYDPLKLISKLGA
KHIKDWEHLEEFRRSLCDVAVSLNNCAYYTQLDDAVWEVHKTAPPGSFVYKSLVKYLSD
KVLFRSLFIDGSSC"
gene 4394..5812
/gene="RNA pol"
/label="RNA pol"
CDS 5796..6599
/gene="MP"
/label="Movement protein"
/note="Movement protein from Tobacco mosaic virus (strain
vulgare). Accession#: P03583"
protein_bind 6674..6798
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 6823..6853
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
CDS 6907..7563
/label="CmR"
/note="chloramphenicol acetyltransferase"
CDS 7908..8210
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(8254..8378)
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 8401..8424
/label="HRV 3C site"
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
CDS 8440..8463
/label="Strep-Tag II"
/note="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
CDS 8500..8523
/label="Strep-Tag II"
/note="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
CDS 8542..8559
/label="6xHis"
/note="6xHis affinity tag"
regulatory 8892..9102
/label="CaMV 35S"
/note="CaMV 35S"
/regulatory_class="terminator"
misc_feature 9172..9196
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS complement(9351..10496)
/label="trfA"
/note="trans-acting replication protein that binds to and
activates oriV"
CDS complement(10798..11589)
/label="KanR"
/note="aminoglycoside phosphotransferase"
rep_origin complement(11801..12436)
/direction=LEFT
/label="oriV"
/note="incP origin of replication"
This page is informational only.