hygII-pSmRYNE(R) vector (V009911)

Basic Vector Information

      • Vector Name:
      • hygII-pSmRYNE(R)
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 13281 bp
      • Type:
      • BiFC expression vector
      • Source/Author:
      • Offenborn JN, Waadt R, Kudla J.

hygII-pSmRYNE(R) vector Vector Map

hygII-pSmRYNE(R)13281 bp600120018002400300036004200480054006000660072007800840090009600102001080011400120001260013200RK2 oriVprokaryotic transposable elementNptIIIRK2 TrfAT-DNA left borderM13 fwdpAg7HptIIpNOSpSUPERdBammRYNEMCS2NosTT-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-pSmRYNE(R) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               13281 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  BiFC expression vector hygII-pSmRYNE(R), complete sequence.
ACCESSION   KR857719
VERSION     .
KEYWORDS    BiFC expression vector.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13281)
  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 13281)
  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 13281)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13281
                     /organism="BiFC expression vector hygII-pSmRYNE(R)"
                     /mol_type="other DNA"
                     /db_xref="taxon:1662161"
     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="AKJ78256.1"
                     /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="AKJ78257.1"
                     /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..9804
                     /label=pSUPERdBam
                     /note="pSUPERdBam"
     promoter        9422..9802
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             9814..10314
                     /codon_start=1
                     /product="mRYNE(R)"
                     /label=mRYNE
                     /protein_id="AKJ78255.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDDYKD
                     DDDK"
     misc_feature    9814..10290
                     /label=mCherry 1-159
                     /note="mCherry 1-159"
     misc_feature    10291..10314
                     /label=FLAG-Tag
                     /note="FLAG-Tag"
     CDS             10291..10314
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    10315..10365
                     /label=MCS2
                     /note="MCS2"
     misc_feature    10375..10639
                     /label=NosT
                     /note="NosT"
     terminator      10385..10637
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(10764..10925)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    10804..10828
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11293..11943
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     oriT            12624..12733
                     /note="incP origin of transfer"
     CDS             12766..13137
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      13264..694
                     /label=oriV
                     /note="incP origin of replication"

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