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phMGFP vector (V011998) Gene synthesis in phMGFP backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V011998 phMGFP In stock, instant shipping

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

Basic Vector Information

phMGFP is a mammalian expression vector encoding Monster Green® Fluorescent Protein, a codon-optimized synthetic variant of Montastrea cavernosa GFP. The 26kDa hMGFP protein offers 20% brighter fluorescence than commercial GFPs, reduced cytotoxicity, and minimized cellular perturbations, making it an ideal choice for reliable, high-level expression in both transient/stable assays.

Vector Name:
phMGFP
Antibiotic Resistance:
Ampicillin
Length:
4707 bp
Type:
Fluorescent Protein Genes & Plasmids
Replication origin:
ori
Source/Author:
Promega
Copy Number:
High copy number
Promoter:
CMV

phMGFP vector Map

Copy Sequence View Map
phMGFP4707 bp600120018002400300036004200CMV enhancer/promoterchimeric intronT7 promoterhMGFPSV40 late polyadenylation signalAmpR

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.

References

  • Afrin R, Zohora US, Uehara H, Watanabe-Nakayama T, Ikai A. Atomic force microscopy for cellular level manipulation: imaging intracellular structures and DNA delivery through a membrane hole. J Mol Recognit. 2009 Sep-Oct;22(5):363-72. doi: 10.1002/jmr.971. PMID: 19623603.

phMGFP vector Sequence

LOCUS       AY218848                4707 bp    DNA     circular SYN 08-MAR-2003
DEFINITION  Monster GFP vector phMGFP, complete sequence.
ACCESSION   AY218848
VERSION     AY218848.1
KEYWORDS    .
SOURCE      Monster GFP vector phMGFP
  ORGANISM  Monster GFP vector phMGFP
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4707)
  AUTHORS   Sundquist,T., Chauvin,F. and Almond,B.
  TITLE     Monster Green Fluorescent Protein phMGFP Vector Technical Bulletin,
            TB320
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4707)
  AUTHORS   Sundquist,T., Chauvin,F. and Almond,B.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JAN-2003) Scientific Communications, Promega
            Corporation, 2800 Woods Hollow Rd, Madison, WI 53711, USA
FEATURES             Location/Qualifiers
     source          1..4707
                     /organism="Monster GFP vector phMGFP"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:220994"
                     /note="green fluorescent protein cloned from Montastraea
                     cavernosa, the great star coral"
     regulatory      1..742
                     /regulatory_class="promoter"
                     /note="CMV enhancer/promoter"
     intron          857..989
                     /note="chimeric intron"
     regulatory      1034..1052
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     CDS             1076..1759
                     /note="hMGFP"
                     /codon_start=1
                     /product="green fluorescent protein"
                     /protein_id="AAO43180.1"
                     /translation="MGVIKPDMKIKLRMEGAVNGHKFVIEGDGKGKPFEGKQTMDLTV
                     IEGAPLPFAYDILTTVFDYGNRVFAKYPKDIPDYFKQTFPEGYSWERSMTYEDQGICI
                     ATNDITMMKGVDDCFVYKIRFDGVNFPANGPVMQRKTLKWEPSTEKMYVRDGVLKGDV
                     NMALLLEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHRIEIVSHDKDYNKVKLYEHAEA
                     HSGLPRQAG"
     misc_feature    1807..2028
                     /note="SV40 late polyadenylation signal"
     CDS             3015..3875
                     /note="AmpR"
                     /codon_start=1
                     /product="B-lactamase"
                     /protein_id="AAO43181.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"