hygII-mRYNE vector (V009912)

Basic Vector Information

      • Vector Name:
      • hygII-mRYNE
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 13050 bp
      • Type:
      • BiFC expression vector
      • Source/Author:
      • Offenborn JN, Waadt R, Kudla J.

hygII-mRYNE vector Vector Map

hygII-mRYNE13050 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600RK2 oriVprokaryotic transposable elementNptIIIRK2 TrfAT-DNA left borderM13 fwdpAg7HptIIpNOSp35SMCSmRYNENosTT-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 ℃.

hygII-mRYNE vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               13050 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  BiFC expression vector hygII-mRYNE, complete sequence.
ACCESSION   KR857708
VERSION     .
KEYWORDS    BiFC expression vector.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13050)
  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 13050)
  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 13050)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13050
                     /organism="BiFC expression vector hygII-mRYNE"
                     /mol_type="other DNA"
                     /db_xref="taxon:1662158"
     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="AKJ78223.1"
                     /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(7023..8048)
                     /codon_start=1
                     /product="hygromycin resistance protein"
                     /label=hygromycin resistance protein
                     /note="HptII"
                     /protein_id="AKJ78224.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGAFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    complement(8077..8364)
                     /label=pNOS
                     /note="pNOS"
     promoter        complement(8102..8281)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    8686..9549
                     /label=p35S
                     /note="p35S"
     promoter        9196..9541
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    9550..9633
                     /label=MCS
                     /note="MCS"
     CDS             9634..10137
                     /codon_start=1
                     /product="mRYNE"
                     /label=mRYNE
                     /protein_id="AKJ78222.1"
                     /translation="MDYKDDDDKVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGE
                     GEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGF
                     KWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERM
                     YPED"
     misc_feature    9637..9660
                     /label=FLAG-Tag
                     /note="FLAG-Tag"
     CDS             9637..9660
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    9661..10134
                     /label=mCherry 2-159
                     /note="mCherry 2-159"
     misc_feature    10144..10408
                     /label=NosT
                     /note="NosT"
     terminator      10154..10406
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(10533..10694)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    10573..10597
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11062..11712
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     oriT            12393..12502
                     /note="incP origin of transfer"
     CDS             12535..12906
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      13033..694
                     /label=oriV
                     /note="incP origin of replication"

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