pMSD1 vector (V004462)

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V004462	pMSD1	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:: pMSD1

Length:: 5602 bp

Type:: Cloning vector

Replication origin:: pBBR1 oriV

Source/Author:: Meng J, Wang H, Liu X, Lin J, Pang X, Lin J.

pMSD1 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.

pMSD1 vector Sequence

LOCUS       40924_32385        5602 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMSD1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5602)
  AUTHORS   Meng J, Wang H, Liu X, Lin J, Pang X, Lin J.
  TITLE     Construction of small plasmid vectors for use in genetic improvement
            of the extremely acidophilic Acidithiobacillus caldus
  JOURNAL   Microbiol. Res. 168 (8), 469-476 (2013)
  PUBMED    23639949
REFERENCE   2  (bases 1 to 5602)
  AUTHORS   Meng J, Wang H, Liu X.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-FEB-2013) State Key Laboratory of Microbial 
            Technology, Shandong University, Shanda Nanlu 27#, Jinan, Shandong 
            250100, China
REFERENCE   3  (bases 1 to 5602)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5602)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol. 
            Res."; date: "2013"; volume: "168"; issue: "8"; pages: "469-476"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-FEB-2013) State Key Laboratory of Microbial Technology, Shandong
            University, Shanda Nanlu 27#, Jinan, Shandong 250100, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3215..3322
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     CDS             3425..4228
                     /codon_start=1
                     /gene="strA"
                     /product="Sm resistance protein A"
                     /label=strA
                     /protein_id="AGT62532.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     gene            3425..4228
                     /gene="strA"
                     /label=strA
     CDS             4228..5064
                     /codon_start=1
                     /gene="strB"
                     /product="Sm resistance protein B"
                     /label=strB
                     /protein_id="AGT62533.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            4228..5064
                     /gene="strB"
                     /label=strB