pDXW-3 vector (V007707)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V007707	pDXW-3	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:: pDXW-3

Antibiotic Resistance:: Kanamycin

Length:: 5067 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Tan Y, Xu D, Li Y, Wang X.

pDXW-3 vector Map

Copy Sequence View Map

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.

pDXW-3 vector Sequence

LOCUS       40924_16390        5067 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDXW-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  AUTHORS   Tan Y, Xu D, Li Y, Wang X.
  TITLE     Construction of a novel sacB-based system for marker-free gene 
            deletion in Corynebacterium glutamicum
  JOURNAL   Plasmid 67 (1), 44-52 (2012)
  PUBMED    22100974
REFERENCE   2  (bases 1 to 5067)
  AUTHORS   Wang X, Tan Y, Xu D, Li Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) State Key Laboratory of Food Science and 
            Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 
            214000, China
REFERENCE   3  (bases 1 to 5067)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5067)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2012"; volume: "67"; issue: "1"; pages: "44-52"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-OCT-2011) State Key Laboratory of Food Science and Technology, 
            Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214000, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    158..203
                     /label=MCS
                     /note="MCS"
     CDS             complement(207..236)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     regulatory      237..282
                     /label=PlacM
                     /note="PlacM"
                     /regulatory_class="promoter"
     CDS             298..1716
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     RBS             complement(1852..1874)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2362..2550
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2655..2797
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3693..4505
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4601..5056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"