pSK104 vector (V003163)

LOCUS       V003163                12392 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003163
VERSION     V003163
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12392)
  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 12392)
  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 12392)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12392)
  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..12392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        85..754
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    766..821
                     /label="TMV Omega"
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             823..4173
                     /codon_start=1
                     /gene="Rep"
                     /product="replicase"
                     /label="Rep"
                     /protein_id="AFX83498.1"
                     /translation="MAYTQTATTSALLDTVRGNNSLVNDLAKRRLYDTAVEEFNARDRR
                     PKVNFSKVISEEQTLIATRAYPEFQITFYNTQNAVHSLAGGLRSLELEYLMMQIPYGSL
                     TYDIGGNFASHLFKGRAYVHCCMPNLDVRDIMRHEGQKDSIELYLSRLERGGKTVPNFQ
                     KEAFDRYAEIPEDAVCHNTFQTCEHQPMQQSGRVYAIALHSIYDIPADEFGAALLRKNV
                     HTCYAAFHFSENLLLEDSCVNLDEINACFSRDGDKLTFSFASESTLNYCHSYSNILKYV
                     CKTYFPASNREVYMKEFLVTRVNTWFCKFSRIDTFLLYKGVAHKSVDSEQFYTAMEDAW
                     HYKKTLAMCNSERILLEDSSSVNYWFPKMRDMVIVPLFDISLETSKRTRKEVLVSKDFV
                     FTVLNHIRTYQAKALTYANVLSFVESIRSRVIINGVTARSEWDVDKSLLQSLSMTFYLH
                     TKLAVLKDDLLISKFSLGSKTVCQHVWDEISLAFGNAFPSVKERLLNRKLIRVAGDALE
                     IRVPDLYVTFHDRLVTEYKASVDMPALDIRKKMEETEVMYNALSELSVLRESDKFDVDV
                     FSQMCQSLEVDPMTAAKVIVAVMSNESGLTLTFERPTEANVALALQDQEKASEGALVVT
                     SREVEEPSMKGSMARGELQLAGLAGDHPESSYSKNEEIESLEQFHMATADSLIRKQMSS
                     IVYTGPIKVQQMKNFIDSLVASLSAAVSNLVKILKDTAAIDLETRQKFGVLDVASRKWL
                     IKPTAKSHAWGVVETHARKYHVALLEYDEQGVVTCDDWRRVAVSSESVVYSDMAKLRTL
                     RRLLRNGEPHVSSAKVVLVDGVPGCGKTKEILSRVNFDEDLILVPGKQAAEMIRRRANS
                     SGIIVATKDNVKTVDSFMMNFGKSTRCQFKRLFIDEGLMLHTGCVNFLVAMSLCEIAYV
                     YGDTQQIPYINRVSGFPYPAHFAKLEVDEVETRRTTLRCPADVTHYLNRRYEGFVMSTS
                     SVKKSVSQEMVGGAAVINPISKPLHGKILTFTQSDKEALLSRGYSDVHTVHEVQGETYS
                     DVSLVRLTPTPVSIIAGDSPHVLVALSRHTCSLKYYTVVMDPLVSIIRDLEKLSSYLLD
                     MYKVDAGTQ"
     gene            823..4173
                     /gene="Rep"
                     /label="Rep"
     CDS             4249..5667
                     /codon_start=1
                     /gene="RNA pol"
                     /product="RNA polymerase"
                     /label="RNA pol"
                     /protein_id="AFX83499.1"
                     /translation="MQFYYDKCLPGNSTMMNNFDAVTMRLTDISLNVKDCILDMSKSVA
                     APKDQIKPLIPMVRTAAEMPRQTGLLENLVAMIKRNFNAPELSGIIDIENTASLVVDKF
                     FDSYLLKEKRKPNKNVSLFSRESLNRWLEKQEQVTIGQLADFDFVDLPAVDQYRHMIKA
                     QPKQKLDTSIQTEYPALQTIVYHQKRSMQYRPLLVSLPGNYWTVLIRADFVFTRKTPAQ
                     IEDFFGDLDSHVPMDVLELDISKYDKSQNEFHCAVEYEIWRRLGFEDFLGEVWKQGHRK
                     TTLKDYTAGIKTCIWYQRKSGDVTTFIGNTVIIAACLASMLPMEKIIKGAFCGDDSLLY
                     FPKGCEFPDVQHSANLMWNFEAKLFKKQYGYFCGRYVIHHDRGCIVYYDPLKLISKLGA
                     KHIKDWEHLEEFRRSLCDVAVSLNNCAYYTQLDDAVWEVHKTAPPGSFVYKSLVKYLSD
                     KVLFRSLFIDGSSC"
     gene            4249..5667
                     /gene="RNA pol"
                     /label="RNA pol"
     CDS             5651..6454
                     /gene="MP"
                     /label="Movement protein"
                     /note="Movement protein from Tobacco mosaic virus (strain
                     vulgare). Accession#: P03583"
     CDS             6517..6594
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags"
     protein_bind    6598..6717
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6747..6777
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6831..7487
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             7832..8134
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(8178..8302)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    8543..8587
                     /note="TobRV/G12a_Ribozyme: aligns with Tobacco ringspot
                     virus satellite RNA and Hammerhead ribozyme G12a"
     misc_feature    8910..8934
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(9089..10234)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     CDS             complement(10536..11327)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(11539..12174)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"
     misc_feature    12252..12276
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     12357..12373
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"