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pDIVA vector (V007992)

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V007992 pDIVA In stock, 1 week for quality controls

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pDIVA
Antibiotic Resistance:
Kanamycin
Length:
4311 bp
Type:
Binary vector
Replication origin:
oriV
Host:
Plants
Source/Author:
Laufer M, Mohammad H, Varrelmann M, Maiss E.
Promoter:
CaMV 35S

pDIVA vector Map

Copy Sequence View Map
pDIVA4311 bp600120018002400300036004200oriVKanRtrfARB T-DNA repeatCaMV 35S promoterHDVagrz35 bp spacerpA35SKS primerLB T-DNA repeatoriV

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pDIVA vector Sequence

LOCUS       40924_14725        4311 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pDIVA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4311)
  AUTHORS   Laufer M, Mohammad H, Varrelmann M, Maiss E.
  TITLE     Construction of infectious full-length cDNA clones of Beet 
            soil-borne mosaic virus and Beet necrotic yellow vein virus A-type 
            by isothermal in vitro recombination analysis of systemic movement, 
            vector transmissions and transreplication properties
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4311)
  AUTHORS   Laufer M, Mohammad H, Varrelmann M, Maiss E.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-AUG-2016) Phytomedicine, Leibniz Universitaet Hannover
            - Institute of Horticultural Production Systems, Herrenhaeuser 
            Strasse 2, Hannover 30419, Germany
REFERENCE   3  (bases 1 to 4311)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4311)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-AUG-2016) Phytomedicine, Leibniz Universitaet Hannover - 
            Institute of Horticultural Production Systems, Herrenhaeuser Strasse
            2, Hannover 30419, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method :: CLC Main Workbench version 6.8.1 Sequencing 
            Technology      :: Sanger dideoxy sequencing ##Assembly-Data-END## 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..622
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     CDS             839..1630
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             1932..3077
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MAQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    complement(3235..3259)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        3412..3754
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    3755..3839
                     /label=HDVagrz
                     /note="HDVagrz"
     misc_feature    3840..3874
                     /label=35 bp spacer
                     /note="35 bp spacer"
     misc_feature    3875..4079
                     /label=pA35S
                     /note="pA35S"
     primer_bind     complement(4109..4125)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(4206..4230)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      4310..4311
                     /label=oriV
                     /note="incP origin of replication"