pTK251 vector (V002622)

LOCUS       V002622                12543 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002622
VERSION     V002622
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12543)
  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 12543)
  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 12543)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12543)
  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..12543
                     /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="AFX83519.1"
                     /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="AFX83520.1"
                     /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"
     CDS             6683..6706
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             6743..6766
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             6800..6817
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             6821..6853
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             6857..6880
                     /label="Xpress(TM) tag"
                     /note="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
     protein_bind    6890..7014
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7039..7069
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             7123..7779
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             8124..8426
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(8470..8594)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    8839..8883
                     /note="TobRV/G12a_Ribozyme: aligns with Tobacco ringspot
                     virus satellite RNA and Hammerhead ribozyme G12a"
     regulatory      8926..9136
                     /label="CaMV 35S"
                     /note="CaMV 35S"
                     /regulatory_class="terminator"
     misc_feature    9206..9230
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(9385..10530)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     CDS             complement(10832..11623)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(11835..12470)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"