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pBBR1MCS2-mamF-Myc vector (V014887)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014887 pBBR1MCS2-mamF-Myc 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:
pBBR1MCS2-mamF-Myc
Antibiotic Resistance:
Kanamycin
Length:
5594 bp
Type:
Protein expression
Replication origin:
pBBR1 oriV
Host:
Broad host
Copy Number:
Low
Promoter:
Lac
Growth Temperature:
37℃

pBBR1MCS2-mamF-Myc vector Map

Copy Sequence View Map
pBBR1MCS2-mamF-Myc5594 bp60012001800240030003600420048005400NeoR/KanRCAP binding sitelac promoterlac operatorM13 revT3 promoterKS primerMagnetosome protein MamFMycSK primerT7 promoterM13 fwdpBBR1 ReppBBR1 oriV

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.

pBBR1MCS2-mamF-Myc vector Sequence

LOCUS       V014887                 5594 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014887
VERSION     V014887
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5594)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             458..1249
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     protein_bind    1657..1678
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1693..1723
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /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     1755..1771
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        1792..1810
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1840..1856
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             1877..2209
                     /gene="mamF"
                     /label="Magnetosome protein MamF"
                     /note="Magnetosome protein MamF from Magnetospirillum
                     gryphiswaldense (strain DSM 6361 / JCM 21280 / NBRC 15271 /
                     MSR-1). Accession#: Q6NE74"
     CDS             2306..2335
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     primer_bind     complement(2340..2356)
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(2389..2407)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2417..2433)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3143..3802)
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(3803..4572)
                     /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"