p35S-GFP vector (V009750)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V009750	p35S-GFP	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

p35S-GFP carries the gene for the green fluorescent protein (GFP) from the bioluminescent jellyfish Aequorea victoria. p35S-GFP was constructed by replacing the GUS coding sequence of pBI221 with a functional GFP gene, thereby placing the GFP gene under the control of the CaMV 35S promoter. Protoplasts were viewed by incident-light fluorescence microscopy 24h after electroporation. 20-60% of the protoplasts emitted an intense green light when illuminated with blue (450-490 nm) light.

Vector Name:: p35S-GFP

Antibiotic Resistance:: Ampicillin

Length:: 4518 bp

Type:: Plant GFP

Replication origin:: ori

Host:: Plants

Source/Author:: Niedz RP, Sussman MR, Satterlee JS.

Promoter:: CaMV35S(long)

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

p35S-GFP 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.

References

Niedz RP, Sussman MR, Satterlee JS. Green fluorescent protein: an in vivo reporter of plant gene expression. Plant Cell Rep. 1995, Apr;14(7):403-6.

Lambardi M, Lachance D, Séguin A, Charest PJ. Evaluation of microprojectile-mediated DNA delivery and reporter genes for genetic transformation of the Mediterranean cypress (Cupressus sempervirens L.). Plant Cell Rep. 1998 Dec;18(3-4):198-202.

p35S-GFP vector Sequence

LOCUS       40924_2622        4518 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p35S-GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4518)
  AUTHORS   Niedz RP, Sussman MR, Satterlee JS.
  TITLE     Green fluorescent protein: an in vivo reporter of plant gene 
            expression
  JOURNAL   Plant Cell Rep. 14 (7), 403-406 (1995)
  PUBMED    24185445
REFERENCE   2  (bases 1 to 4518)
  AUTHORS   Kitts PA.
  TITLE     CLONTECH Vectors On Disk version 1.3
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4518)
  AUTHORS   Kitts PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JUN-1995) Paul A. Kitts, CLONTECH Laboratories, Inc., 
            4030 Fabian Way, Palo Alto, CA 94303, USA
REFERENCE   4  (bases 1 to 4518)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4518)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell 
            Rep."; date: "1995"; volume: "14"; issue: "7"; pages: "403-406"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (05-JUN-1995) Paul A. Kitts, CLONTECH Laboratories, Inc., 4030 
            Fabian Way, Palo Alto, CA 94303, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     This vector can be obtained from CLONTECH Laboratories, Inc., 4030 
            Fabian Way, Palo Alto, CA 94303, USA. To place an order call (415) 
            424-8222 or (800) 662-2566, extension 1. International customers, 
            please contact your local distributor. For technical information, 
            call (415) 424-8222 or (800) 662-2566, extension 3. This sequence 
            has been compiled from information in the sequence databases, 
            published literature and other sources, together with partial 
            sequences obtained by CLONTECH. If you suspect there is an error in 
            this sequence, please contact CLONTECH's Technical Service 
            Department at (415) 424-8222 or (800) 662-2566, extension 3 or 
            E-mail TECH@CLONTECH.COM.
FEATURES             Location/Qualifiers
     source          1..4518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        512..857
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             896..1609
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1629..1881
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1890..1906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2380..2484
                     /label=AmpR promoter
     CDS             2485..3342
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3516..4104
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4392..4413
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4428..4458
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4466..4482
                     /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     4490..4506
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"