MSP955 vector (V016994)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016994 MSP955 In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
MSP955
Antibiotic Resistance:
Ampicillin
Length:
10017 bp
Type:
Protein expression
Replication origin:
pSC101 ori
Host:
E. coli
Promoter:
T7
Growth Strain(s):
DH5a
Growth Temperature:
30℃

MSP955 vector Map

MSP95510017 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000Rep101pSC101 oriT7 promoterpIIILuxALuxBT7 RNA polymeraseTEV siteT7 lysozymerrnB T1 terminatorAmpRAmpR promoter

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.

MSP955 vector Sequence

LOCUS       V016994                10017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016994
VERSION     V016994
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10017)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..951)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     rep_origin      complement(999..1221)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        1785..1803
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1985..3256
                     /label="M13 gene III"
                     /note="pIII"
     CDS             3259..4338
                     /label="LuxA"
                     /note="LuxA luciferase subunit"
     CDS             4402..5379
                     /label="LuxB"
                     /note="LuxB luciferase subunit"
     CDS             complement(5482..8127)
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     CDS             complement(8161..8181)
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             complement(8212..8664)
                     /label="T7 lysozyme"
                     /note="lysozyme from bacteriophage T7"
     terminator      complement(8849..8895)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(8949..9806)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9807..9911)
                     /label="AmpR promoter"