Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008094 | pCX4pur | 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:
- pCX4pur
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6076 bp
- Type:
- Retroviral vector
- Replication origin:
- ori
- Source/Author:
- Akagi T, Sasai K, Hanafusa H.
pCX4pur vector 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.
pCX4pur vector Sequence
LOCUS 40924_13965 6076 bp DNA circular SYN 17-DEC-2018
DEFINITION Retroviral vector pCX4pur DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6076)
AUTHORS Akagi T, Sasai K, Hanafusa H.
TITLE Refractory nature of normal human diploid fibroblasts with respect
to oncogene-mediated transformation
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 100 (23), 13567-13572 (2003)
PUBMED 14597713
REFERENCE 2 (bases 1 to 6076)
AUTHORS Akagi T.
TITLE Direct Submission
JOURNAL Submitted (10-JUN-2002) Tsuyoshi Akagi, Osaka Bioscience Institute,
Molecular Oncology; Furuedai 6-2-4, Suita, Osaka 565-0874, Japan
(E-mail:takagi@obi.or.jp, Tel:81-6-6872-4834, Fax:81-6-6871-7521)
REFERENCE 3 (bases 1 to 6076)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6076)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
Acad. Sci. U.S.A."; date: "2003"; volume: "100"; issue: "23"; pages:
"13567-13572"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(10-JUN-2002) Tsuyoshi Akagi, Osaka Bioscience Institute, Molecular
Oncology"; volume: " Furuedai 6-2-4, Suita, Osaka 565-0874, Japan
(E-mail:takagi@obi.or.jp, Tel:81-6-6872-4834, Fax"; pages:
"81-6-6871-7521"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6076
/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 1242..1658
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
misc_feature 1668..2042
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
misc_feature 2044..2106
/label=multiple cloning site
/note="multiple cloning site"
misc_feature 2154..2715
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2718..3314
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
LTR 3371..3962
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"
rep_origin complement(4263..4851)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5018..5875)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5876..5980)
/label=AmpR promoter