DsRed-Rab5 WT vector (V001617)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V001617	DsRed-Rab5 WT	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: DsRed-Rab5 WT

Antibiotic Resistance:: Kanamycin

Length:: 5369 bp

Type:: Mammalian Expression

Replication origin:: ori

Copy Number:: High Copy

Cloning Method:: Restriction Enzyme

DsRed-Rab5 WT vector Map

Copy Sequence View Map

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.

DsRed-Rab5 WT vector Sequence

LOCUS       V001617                 5369 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001617
VERSION     V001617
KEYWORDS    DsRed-Rab5 WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5369)
  AUTHORS   Sharma DK, Choudhury A, Singh RD, Wheatley CL, Marks DL, Pagano RE
  TITLE     Glycosphingolipids internalized via caveolar-related endocytosis
            rapidly merge with the clathrin pathway in early endosomes and form
            microdomains for recycling.
  JOURNAL   J Biol Chem. 2003 Feb 28. 278(9):7564-72.
   PUBMED   12482757
REFERENCE   2  (bases 1 to 5369)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5369)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
            Chem. 2003 Feb 28. 278(9):7564-72."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             568..1242
                     /label="DsRed2"
                     /note="improved tetrameric variant of DsRed fluorescent
                     protein"
     regulatory      1249..1258
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1261..1287
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1291..1932
                     /gene="RAB5A"
                     /label="Ras-related protein Rab-5A"
                     /note="Ras-related protein Rab-5A from Homo sapiens.
                     Accession#: P20339"
     misc_feature    1936..1988
                     /label="MCS"
                     /note="multiple cloning site"
     polyA_signal    2112..2233
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2240..2695)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2722..2826
                     /label="AmpR promoter"
     promoter        2828..3185
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3220..4011
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     primer_bind     complement(4202..4221)
                     /label="TK-pA-R"
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4246..4293
                     /label="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4622..5210
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5325..5369
                     /label="CMV"
                     /note="Human cytomegalovirus immediate early
                     enhancer/promoter"