phMGFP vector (V011998)

Price Information

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V011998 phMGFP In stock, instant shipping

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

Basic Vector Information

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

phMGFP4707 bp600120018002400300036004200CMV enhancerCMV promoterchimeric intronT7 promotermc4T3 promoterSV40 poly(A) signalf1 oriAmpR promoterAmpRori

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.

phMGFP vector Sequence

LOCUS       40924_24732        4707 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Monster GFP vector phMGFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4707)
  AUTHORS   Sundquist T, Chauvin F, 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, Almond B.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JAN-2003) Scientific Communications, Promega 
            Corporation, 2800 Woods Hollow Rd, Madison, WI 53711, USA
REFERENCE   3  (bases 1 to 4707)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4707)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JAN-2003) Scientific Communications, Promega Corporation, 2800 
            Woods Hollow Rd, Madison, WI 53711, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1076..1753
                     /codon_start=1
                     /label=mc4
                     /note="mc4 is a basic (constitutively fluorescent)
                     green/yellow fluorescent protein published in 2003, derived
                     from Montastraea cavernosa."
                     /translation="MGVIKPDMKIKLRMEGAVNGHKFVIEGDGKGKPFEGKQTMDLTVI
                     EGAPLPFAYDILTTVFDYGNRVFAKYPKDIPDYFKQTFPEGYSWERSMTYEDQGICIAT
                     NDITMMKGVDDCFVYKIRFDGVNFPANGPVMQRKTLKWEPSTEKMYVRDGVLKGDVNMA
                     LLLEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHRIEIVSHDKDYNKVKLYEHAEAHSGL
                     PRQA"
     promoter        complement(1781..1798)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    complement(1816..1937)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2123..2578
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2910..3014
                     /label=AmpR promoter
     CDS             3015..3872
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4046..4634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"