pMW82 vector (V004383)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004383	pMW82	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pMW82

Antibiotic Resistance:: Ampicillin

Length:: 5129 bp

Type:: Promoter-trap vector

Replication origin:: ori

Source/Author:: Bumann D.

pMW82 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.

pMW82 vector Sequence

LOCUS       40924_32818        5129 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter-trap vector pMW82, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5129)
  AUTHORS   Bumann D.
  TITLE     In vivo visualization of bacterial colonization, antigen expression,
            and specific T-cell induction following oral administration of live 
            recombinant Salmonella enterica serovar Typhimurium
  JOURNAL   Infect. Immun. 69 (7), 4618-4626 (2001)
  PUBMED    11402006
REFERENCE   2  (bases 1 to 5129)
  AUTHORS   Rollenhagen C, Sorensen M, Rizos K, Hurvitz R, Bumann D.
  TITLE     Antigen selection based on expression levels during infection 
            facilitates vaccine development for an intracellular pathogen
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 101 (23), 8739-8744 (2004)
  PUBMED    15173591
REFERENCE   3  (bases 1 to 5129)
  AUTHORS   Wendland M, Bumann D.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JAN-2007) Molecular Biology, Max-Planck-Institute for 
            Infection Biology, Chartieplatz 1, Berlin D-10117, Germany
REFERENCE   4  (bases 1 to 5129)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5129)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Infect. 
            Immun."; date: "2001"; volume: "69"; issue: "7"; pages: "4618-4626"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2004"; volume: "101"; issue: "23"; pages:
            "8739-8744"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (16-JAN-2007) Molecular Biology, Max-Planck-Institute for Infection 
            Biology, Chartieplatz 1, Berlin D-10117, Germany"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             32..745
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1038..1124
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1216..1243
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1262..1353
                     /label=AmpR promoter
     CDS             1354..2211
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2385..2973
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3159..3299)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3404..3592)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"