Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V010446 | pLNCX2 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pLNCX2 is a retroviral vector designed for stable gene expression in mammalian cells. It contains a neomycin resistance gene (neo) for antibiotic selection and a multiple cloning site (MCS) to insert target genes. The expression of inserted genes is driven by the retroviral LTR promoter or an optional internal promoter. This plasmid enables efficient gene delivery via retroviral transduction and integrates into the host genome for long-term expression. It's widely used in functional gene studies, generating stable cell lines, and investigating gene regulation. The LTRs flanking the cassette ensure proper viral packaging when co-transfected with helper plasmids.
- Vector Name:
- pLNCX2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6132 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- CMV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLNCX2 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.
References
- Takekoshi T, Fang L, Paragh G, Hwang ST. CCR7-expressing B16 melanoma cells downregulate interferon-γ-mediated inflammation and increase lymphangiogenesis in the tumor microenvironment. Oncogenesis. 2012 May 7;1(5):e9. doi: 10.1038/oncsis.2012.9. PMID: 23552640; PMCID: PMC3412639.
pLNCX2 vector Sequence
LOCUS 40924_28536 6132 bp DNA circular SYN 01-JAN-1980
DEFINITION Retroviral vector for transient or stable constitutive expression of
a gene.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6132)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6132)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6132
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 2..589
/label=5' LTR
/note="long terminal repeat from Moloney murine sarcoma
virus"
misc_feature 652..851
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1052..1468
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
CDS 1512..2303
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
enhancer 2377..2680
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2681..2884
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 2925..2994
/label=MCS
/note="MCS"
/note="multiple cloning site"
LTR 3034..3627
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 3838..3978
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4164..4752)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4926..5783)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5784..5888)
/label=AmpR promoter