pSKI074 vector (V003154)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V003154	pSKI074	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pSKI074

Antibiotic Resistance:: Ampicillin

Length:: 10450 bp

Type:: Activation tagging vector

Replication origin:: oriV

Source/Author:: Weigel D, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK, Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Wang Z, Xia Y, Dixon RA, Harrison MJ, Lamb CJ, Yanofs

Promoter:: MAS

pSKI074 vector Vector Map

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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.

pSKI074 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_40587       10450 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Activation tagging vector pSKI074, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10450)
  AUTHORS   Weigel D, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK, 
            Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM, 
            Nguyen JT, Sato S, Wang Z, Xia Y, Dixon RA, Harrison MJ, Lamb CJ, 
            Yanofsky MF, Chory J.
  TITLE     Activation tagging in Arabidopsis
  JOURNAL   Plant Physiol. 122 (4), 1003-1013 (2000)
  PUBMED    10759496
REFERENCE   2  (bases 1 to 10450)
  AUTHORS   Weigel D, Blazquez MA, Borevitz J, Christensen SK, Fankhauser C, 
            Ferrandiz C, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Xia Y, 
            Wang Z, Dixon RA, Harrison MJ, Lamb CJ, Yanofsky MF, Chory J.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-DEC-1999) Plant Biology Laboratory, The Salk Institute
            for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 
            92037, USA
REFERENCE   3  (bases 1 to 10450)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10450)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol."; date: "2000"; volume: "122"; issue: "4"; pages: 
            "1003-1013"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-DEC-1999) Plant Biology Laboratory, The Salk Institute for 
            Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(13..31)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(52..68)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(76..92)
                     /label=lac operator
                     /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)."
     promoter        complement(100..130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(145..166)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(454..1042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1216..2073)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2074..2178)
                     /label=AmpR promoter
     rep_origin      complement(2207..2662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     misc_feature    2663..2829
                     /label=derived from pUC19
                     /note="derived from pUC19"
     primer_bind     2807..2823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2830..2848
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2881..2897
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2894..4178
                     /label=from cauliflower mosaic virus 35S gene promoter
                     /note="from cauliflower mosaic virus 35S gene promoter"
                     /regulatory_class="enhancer"
     misc_feature    4270..4294
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     oriT            complement(5497..5606)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      6401..7113
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7700..7724
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     primer_bind     complement(8045..8061)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8362..8741
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             8771..9562
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      9746..10430
                     /label=from mannopine synthase
                     /note="from mannopine synthase"
                     /regulatory_class="terminator"