pCXbsr vector (V008093)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V008093	pCXbsr	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

Vector Name:: pCXbsr

Antibiotic Resistance:: Ampicillin

Length:: 5883 bp

Type:: Retroviral vector

Replication origin:: ori

Source/Author:: Akagi T, Shishido T, Murata K, Hanafusa H.

pCXbsr 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.

pCXbsr vector Sequence

LOCUS       V008093                 5883 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008093
VERSION     V008093
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5883)
  AUTHORS   Akagi T, Shishido T, Murata K, Hanafusa H.
  TITLE     v-Crk activates the phosphoinositide 3-kinase/AKT pathway in
            transformation
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 97 (13), 7290-7295 (2000)
   PUBMED   10852971
REFERENCE   2  (bases 1 to 5883)
  AUTHORS   Akagi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-APR-2000) Tsuyoshi Akagi, Osaka Bioscience Institute,
            The First Department; 6-2-4 Furuedai, Suita, Osaka 565-0874, Japan
            (E-mail:takagi@obisun1.obi.or.jp, Tel:81-6-6872-4850,
            Fax:81-6-6871-6686)
REFERENCE   3  (bases 1 to 5883)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5883)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2000"; volume: "97"; issue: "13"; pages:
            "7290-7295"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (17-APR-2000) Tsuyoshi Akagi, Osaka Bioscience Institute, The First
            Department"; volume: " 6-2-4 Furuedai, Suita, Osaka 565-0874, Japan
            (E-mail:takagi@obisun1.obi.or.jp, Tel:81-6-6872-4850, Fax"; pages:
            "81-6-6871-6686"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..467
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   467..762
                     /label="R-U5 region of MuLV 5'LTR"
                     /note="R-U5 region of MuLV 5'LTR"
     misc_feature    826..1183
                     /label="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1248..1664
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1674..2048
                     /label="pol region"
                     /note="Moloney murine leukemia virus (MMLV) pol region
                     containing the splice acceptor site"
     misc_feature    2052..2136
                     /label="multiple cloning site"
                     /note="multiple cloning site"
     misc_feature    2185..2765
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             2765..3184
                     /gene="bsr"
                     /label="Blasticidin-S deaminase"
                     /note="Blasticidin-S deaminase from Bacillus cereus.
                     Accession#: P33967"
     LTR             3240..3833
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(4070..4658)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4825..5682)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5683..5787)
                     /label="AmpR promoter"