D3cpv-C vector (V010027)

Basic Vector Information

      • Vector Name:
      • D3cpv-C
      • Antibiotic Resistance:
      • Spectinomycin/Streptomycin
      • Length:
      • 12828 bp
      • Type:
      • Binary expression vector
      • Source/Author:
      • Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, Kudla J, Schumacher K.

D3cpv-C vector Vector Map

D3cpv-C12828 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600RB T-DNA repeatCAP binding sitelac promoterlac operatorM13 revAtUBQ10multiple cloning siteCFPVN173E9 terminatorM13 fwdMAS terminatorNeoR/KanRMAS promoterLB T-DNA repeatSmRoribompVS1 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 ℃.

D3cpv-C vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_565       12828 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary expression vector D3cpv-C, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12828)
  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 12828)
  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 12828)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12828)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J."; 
            date: "2012"; volume: "69"; issue: "1"; pages: "181-192"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-OCT-2011) Department of Developmental Biology, Centre for 
            Organismal Studies Heidelberg, Im Neuenheimer Feld 230, Heidelberg 
            69120, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(114..138)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    303..324
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        339..369
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    377..393
                     /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     401..417
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      443..1047
                     /label=AtUBQ10
                     /note="AtUBQ10"
                     /regulatory_class="promoter"
     misc_feature    1077..1130
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             1154..1837
                     /label=CFP
                     /note="cyan variant of GFP"
     CDS             2594..3112
                     /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"
     terminator      3131..3772
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     primer_bind     complement(3785..3801)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(4383..4635)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(4834..5625)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(5655..6035)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    complement(6407..6431)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6959..7747
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     rep_origin      7994..8582
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(8768..8908)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9252..9446)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9515..10579)
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11016..11642)
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"

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