pDendra2-C vector (V012055)

Price Information

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V012055 pDendra2-C 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:
pDendra2-C
Antibiotic Resistance:
Kanamycin
Length:
4716 bp
Type:
Fluorescent Protein Genes & Plasmids
Replication origin:
ori
Source/Author:
Clontech
Selection Marker:
Neomycin/G418(Geneticin)
Copy Number:
High copy number
Promoter:
CMV

pDendra2-C vector Map

pDendra2-C4716 bp600120018002400300036004200CMV enhancerCMV promoterDendra2MCSstop codonsSV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRHSV TK poly(A) signalori

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.

pDendra2-C vector Sequence

LOCUS       pDendra2-C.        4716 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for fusing Dendra2 to the N-terminus of a partner protein.
ACCESSION   .
VERSION     .
KEYWORDS    pDendra2-C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4716)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4716)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             613..1302
                     /label=Dendra2
                     /note="monomeric photoswitchable green-to-red fluorescent 
                     protein from Dendronephthya"
     misc_feature    1328..1380
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1389..1399
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    1504..1625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1632..2087)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2114..2218
                     /label=AmpR promoter
     promoter        2220..2577
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2612..3403
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3638..3685
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4014..4602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"