mCherry-MS2-6×His vector (V014551)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V014551	mCherry-MS2-6×His	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:: mCherry-MS2-6×His

Antibiotic Resistance:: Ampicillin

Length:: 6017 bp

Type:: Gene expression

Replication origin:: ori

Host:: Mammalian cells

Promoter:: T7

Growth Strain(s):: DH5a

Growth Temperature:: 37℃

mCherry-MS2-6×His 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.

mCherry-MS2-6×His vector Sequence

LOCUS       V014551                 6017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014551
VERSION     V014551
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6017)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             750..776
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             777..788
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             1167..1184
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             1185..1892
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     intron          1972..2544
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     CDS             2548..2670
                     /gene="HBB"
                     /label="Hemoglobin subunit beta"
                     /note="Hemoglobin subunit beta from Colobus guereza.
                     Accession#: Q9TT33"
     polyA_signal    2741..2796
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     promoter        complement(3151..3169)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3176..3192)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3334..3789
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3815..3919
                     /label="AmpR promoter"
     misc_feature    4104..4483
                     /label="ISS"
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of
                     5'-AACGTT-3' (Sato et al., 1996)"
     rep_origin      4952..5540
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    5828..5849
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5864..5894
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    5902..5918
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5926..5942
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        5963..5981
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"