pLMB509 vector (V004974)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004974	pLMB509	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:: pLMB509

Antibiotic Resistance:: Gentamycin

Length:: 6879 bp

Type:: Cloning vector

Replication origin:: pBBR1 oriV

Source/Author:: Tett AJ, Rudder SJ, Bourdes A, Karunakaran R, Poole PS.

Promoter:: Pc

pLMB509 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.

pLMB509 vector Sequence

LOCUS       40924_28481        6879 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLMB509, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  AUTHORS   Tett AJ, Rudder SJ, Bourdes A, Karunakaran R, Poole PS.
  TITLE     Regulatable vectors for environmental gene expression in 
            alphaproteobacteria
  JOURNAL   Appl. Environ. Microbiol. 78 (19), 7137-7140 (2012)
  PUBMED    22820336
REFERENCE   2  (bases 1 to 6879)
  AUTHORS   Tett A, Rudder S, Bourdes A, Poole P.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-APR-2012) Molecular Microbiology, John Innes Centre, 
            Colney Lane, Norwich NR47UH, UK
REFERENCE   3  (bases 1 to 6879)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6879)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2012"; volume: "78"; issue: "19"; 
            pages: "7137-7140"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-APR-2012) Molecular Microbiology, John Innes Centre, Colney 
            Lane, Norwich NR47UH, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..884
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     CDS             891..908
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      942..1028
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(1720..2379)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(2380..3149)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3373..4377
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /label=mob
                     /note="mobilisation protein"
                     /protein_id="AFR24685.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLAAL"
     gene            3373..4377
                     /gene="mob"
                     /label=mob
     CDS             complement(4435..4965)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(5154..5182)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     regulatory      complement(5395..6879)
                     /gene="tauA_tauR"
                     /regulatory_class="promoter"
     gene            complement(5395..6879)
                     /gene="tauA_tauR"
                     /label=tauA_tauR