NES-YC3.6 vector (V009836)

Basic Vector Information

      • Vector Name:
      • NES-YC3.6
      • Antibiotic Resistance:
      • Spectinomycin/Streptomycin
      • Length:
      • 12872 bp
      • Type:
      • Binary expression vector
      • Source/Author:
      • Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, Kudla J, Schumacher K.

NES-YC3.6 vector Vector Map

NES-YC3.612872 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600right borderCAP binding sitelac promoterlac operatorM13 revAtUBQ10NES-YC3.6rbcsM13 fwdmasnptIImasleft borderaadAoribompVS1 oriVpVS1 RepApVS1 StaA

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 ℃.

NES-YC3.6 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               12872 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary expression vector NES-YC3.6, complete sequence.
ACCESSION   JN903572
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12872)
  AUTHORS   Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, 
            Kudla J, Schumacher K.
  TITLE     FRET-based genetically encoded sensors allow high-resolution live 
            cell imaging of Ca(2) dynamics
  JOURNAL   Plant J. 69 (1), 181-192 (2012)
  PUBMED    21910770
REFERENCE   2  (bases 1 to 12872)
  AUTHORS   Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, 
            Kudla J, Schumacher K.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-OCT-2011) Department of Developmental Biology, Centre 
            for Organismal Studies Heidelberg, Im Neuenheimer Feld 230, 
            Heidelberg 69120, Germany
REFERENCE   3  (bases 1 to 12872)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12872
                     /organism="Binary expression vector NES-YC3.6"
                     /mol_type="other DNA"
                     /db_xref="taxon:1154702"
     misc_feature    1..194
                     /label=right border
                     /note="right border"
     misc_feature    113..137
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    302..323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        338..368
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    376..392
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     400..416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      444..1088
                     /regulatory_class="promoter"
                     /note="AtUBQ10"
     CDS             1114..3159
                     /codon_start=1
                     /product="NES-YC3.6"
                     /label=NES-YC3.6
                     /protein_id="AFC41195.1"
                     /translation="MLQNELALKLAGLDINKTGGVDSSAAATMVSKGEELFTGVVPILV
                     ELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYP
                     DHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDG
                     NILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPV
                     LLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAARMHDQLTEEQIAEFKEAFSLFDKDG
                     DGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIYFPEFLTMMARKMKDTDSE
                     EEIREAFRVFDKDGNGYISAAQLRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEF
                     VQMMTAKGGKRRWKKNFIAVSAANRFKKISSSGALELMDGGVQLADHYQQNTPIGDGPV
                     LLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELF
                     TGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYG
                     LQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELK
                     GIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIE"
     misc_feature    1114..1174
                     /label=NES
                     /note="NES"
     regulatory      1192..1201
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2338..2415
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2638..3156
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     regulatory      3174..3820
                     /regulatory_class="terminator"
                     /note="rbcs"
     terminator      3175..3816
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     primer_bind     complement(3829..3845)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(4009..4700)
                     /regulatory_class="terminator"
                     /note="mas"
     terminator      4427..4679
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(4875..5669)
                     /codon_start=1
                     /product="nptII"
                     /label=nptII
                     /protein_id="AFC41196.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(5699..6383)
                     /regulatory_class="promoter"
                     /note="mas"
     promoter        complement(5699..6079)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    6384..6712
                     /label=left border
                     /note="left border"
     misc_feature    6451..6475
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6862..7794
                     /codon_start=1
                     /product="aadA"
                     /label=aadA
                     /protein_id="AFC41197.1"
                     /translation="MTCFFGVQSMPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKL
                     NIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVR
                     LDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRND
                     ILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNS
                     PPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQ
                     EEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             7003..7794
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      8038..8626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8812..8952
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      9296..9490
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9556..10623)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     CDS             complement(11057..11686)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"

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