Basic Vector Information
- Vector Name:
- pMW390
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 13180 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)
pMW390 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 ℃.
pMW390 vector Sequence
LOCUS V004386 13180 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V004386 VERSION V004386 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 13180) 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 13180) 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 13180) TITLE Direct Submission REFERENCE 4 (bases 1 to 13180) 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..13180 /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="AFX83540.1" /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="AFX83541.1" /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 6668..6787 /label="attR1" /note="recombination site for the Gateway(R) LR reaction" promoter 6817..6847 /label="lac UV5 promoter" /note="E. coli lac promoter with an 'up' mutation" CDS 6901..7557 /label="CmR" /note="chloramphenicol acetyltransferase" CDS 7902..8204 /label="ccdB" /note="CcdB, a bacterial toxin that poisons DNA gyrase" protein_bind complement(8248..8372) /label="attR2" /note="recombination site for the Gateway(R) LR reaction" CDS 8380..9093 /label="EYFP" /note="enhanced YFP" CDS 9139..9162 /label="FLAG" /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 9166..9189 /label="FLAG" /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 9193..9216 /label="FLAG" /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" misc_feature 9476..9520 /note="TobRV/G12a_Ribozyme: aligns with Tobacco ringspot virus satellite RNA and Hammerhead ribozyme G12a" regulatory 9563..9773 /label="CaMV 35S" /note="CaMV 35S" /regulatory_class="terminator" misc_feature 9843..9867 /label="RB T-DNA repeat" /note="right border repeat from nopaline C58 T-DNA" CDS complement(10022..11167) /label="trfA" /note="trans-acting replication protein that binds to and activates oriV" CDS complement(11469..12260) /label="KanR" /note="aminoglycoside phosphotransferase" rep_origin complement(12472..13107) /direction=LEFT /label="oriV" /note="incP origin of replication"
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