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kanII-mRYCE vector (V009901#)

Basic Vector Information

Basic Vector Information
Vector Name kanII-mRYCE Antibiotic Resistance Kanamycin
Length 12592 bp Type BiFC expression vector
Source Offenborn JN, Waadt R, Kudla J.

kanII-mRYCE vector Vector Map

RK2 oriV prokaryotic transposable element kanamycin resistance protein RK2 TrfA T-DNA left border M13 fwd pAg7 NptII pNOS p35S MCS mRYCE NosT T-DNA right border TetR incP origin of transfer traJ

kanII-mRYCE vector Sequence

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LOCUS       Exported               12592 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  BiFC expression vector kanII-mRYCE, complete sequence.
ACCESSION   KR857710
VERSION     .
KEYWORDS    BiFC expression vector.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12592)
  AUTHORS   Offenborn JN, Waadt R, Kudla J.
  TITLE     Visualization and translocation of ternary 
            Calcineurin-A/Calcineurin-B/Calmodulin-2 protein complexes by 
            dual-color trimolecular fluorescence complementation
  JOURNAL   New Phytol. (2015) In press
  PUBMED    25919910
REFERENCE   2  (bases 1 to 12592)
  AUTHORS   Offenborn JN, Waadt R, Kudla J.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2015) University of Muenster, Institute for 
            Biology and Plant Biotechnology, Schlossplatz 7, Muenster 48149, 
            Germany
REFERENCE   3  (bases 1 to 12592)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12592
                     /organism="BiFC expression vector kanII-mRYCE"
                     /mol_type="other DNA"
                     /db_xref="taxon:1662164"
     misc_feature    1..618
                     /label=RK2 oriV
                     /note="RK2 oriV"
     mobile_element  1317..2084
                     /mobile_element_type="insertion sequence:IS1"
                     /note="prokaryotic transposable element"
     CDS             2276..3070
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /note="NptIII"
                     /protein_id="AKJ78229.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    3079..4560
                     /label=RK2 TrfA
                     /note="RK2 TrfA"
     CDS             3369..4517
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    6043..6190
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    6113..6137
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     6755..6771
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(6784..6995)
                     /label=pAg7
                     /note="pAg7"
     terminator      6784..6984
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     CDS             complement(7031..7834)
                     /codon_start=1
                     /product="neomycin phosphotransferase II"
                     /label=neomycin phosphotransferase II
                     /note="NptII"
                     /protein_id="AKJ78230.1"
                     /translation="MGAWIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(7031..7822)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    complement(7859..8146)
                     /label=pNOS
                     /note="pNOS"
     promoter        complement(7884..8063)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    8468..9331
                     /label=p35S
                     /note="p35S"
     promoter        8978..9323
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    9332..9415
                     /label=MCS
                     /note="MCS"
     CDS             9416..9679
                     /codon_start=1
                     /product="mRYCE"
                     /label=mRYCE
                     /protein_id="AKJ78228.1"
                     /translation="MYPYDVPDYAGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQL
                     PGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK"
     misc_feature    9419..9445
                     /label=HA-Tag
                     /note="HA-Tag"
     CDS             9419..9445
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    9446..9676
                     /label=mCherry 160-236
                     /note="mCherry 160-236"
     misc_feature    9686..9950
                     /label=NosT
                     /note="NosT"
     terminator      9696..9948
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(10075..10236)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    10115..10139
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10604..11254
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     oriT            11935..12044
                     /note="incP origin of transfer"
     CDS             12077..12448
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      12575..694
                     /label=oriV
                     /note="incP origin of replication"

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