Cart 2

kanII-mRYCE(R) vector (V009902#)

Basic Vector Information

Basic Vector Information
Vector Name kanII-mRYCE(R) Antibiotic Resistance Kanamycin
Length 12565 bp Type BiFC expression vector
Source Offenborn JN, Waadt R, Kudla J.

kanII-mRYCE(R) vector Vector Map

kanII-mRYCE(R)12565 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000RK2 oriVprokaryotic transposable elementkanamycin resistance proteinRK2 TrfAT-DNA left borderM13 fwdpAg7NptIIpNOSp35SmRYCEMCS2NosTT-DNA right borderTetRincP origin of transfertraJ

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 ℃.

kanII-mRYCE(R) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               12565 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  BiFC expression vector kanII-mRYCE(R), complete sequence.
ACCESSION   KR857711
VERSION     .
KEYWORDS    BiFC expression vector.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12565)
  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 12565)
  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 12565)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12565
                     /organism="BiFC expression vector kanII-mRYCE(R)"
                     /mol_type="other DNA"
                     /db_xref="taxon:1662165"
     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="AKJ78232.1"
                     /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="AKJ78233.1"
                     /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"
     CDS             9338..9598
                     /codon_start=1
                     /product="mRYCE(R)"
                     /label=mRYCE
                     /protein_id="AKJ78231.1"
                     /translation="MGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIK
                     LDITSHNEDYTIVEQYERAEGRHSTGGMDELYKYPYDVPDYA"
     misc_feature    9341..9571
                     /label=mCherry 160-236
                     /note="mCherry 160-236"
     misc_feature    9572..9598
                     /label=HA-Tag
                     /note="HA-Tag"
     CDS             9572..9598
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    9599..9649
                     /label=MCS2
                     /note="MCS2"
     misc_feature    9659..9923
                     /label=NosT
                     /note="NosT"
     terminator      9669..9921
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(10048..10209)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    10088..10112
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10577..11227
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     oriT            11908..12017
                     /note="incP origin of transfer"
     CDS             12050..12421
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      12548..694
                     /label=oriV
                     /note="incP origin of replication"

This page is informational only.