Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V007112 | pLTR-RD114A | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pLTR-RD114A is a plasmid encoding the RD114 envelope glycoprotein, utilised for lentiviral vector pseudotyping. It facilitates transduction via the ASCT2 receptor for use in gene therapy research.
- Vector Name:
- pLTR-RD114A
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6258 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- SV40
- Cloning Method:
- Restriction Enzyme
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pLTR-RD114A 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
- Colamartino ABL, Lemieux W, Bifsha P, Nicoletti S, Chakravarti N, Sanz J, Roméro H, Selleri S, Béland K, Guiot M, Tremblay-Laganière C, Dicaire R, Barreiro L, Lee DA, Verhoeyen E, Haddad E. Efficient and Robust NK-Cell Transduction With Baboon Envelope Pseudotyped Lentivector. Front Immunol. 2019 Dec 16;10:2873. doi: 10.3389/fimmu.2019.02873. PMID: 31921138; PMCID: PMC6927467.
pLTR-RD114A vector Sequence
LOCUS Exported 6258 bp DNA circular SYN 06-AUG-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6258)
AUTHORS Zhang XY, La Russa VF, Reiser J
TITLE Transduction of bone-marrow-derived mesenchymal stem cells by using
lentivirus vectors pseudotyped with modified RD114 envelope
glycoproteins.
JOURNAL J Virol. 2004 Feb . 78(3):1219-29.
PUBMED 14722277
REFERENCE 2 (bases 1 to 6258)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6258)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6258)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Virol.
2004 Feb . 78(3):1219-29."
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6258
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(28..6258,1..27)
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 209..743
/label=HIV-1 3' LTR
CDS 1220..2812
/codon_start=1
/label=RD114 Env
/translation="MKLPTGMVILCSLIIVRAGFDDPRKAIALVQKQHGKPCECSGGQV
SEAPPNSIQQVTCPGKTAYLMTNQKWKCRVTPKNLTPSGGELQNCPCNTFQDSMHSSCY
TEYRQCRANNKTYYTATLLKIRSGSLNEVQILQNPNQLLQSPCRGSINQPVCWSATAPI
HISDGGGPLDTKRVWTVQKRLEQIHKAMHPELQYHPLALPKVRDDLSLDARTFDILNTT
FRLLQMSNFSLAQDCWLCLKLGTPTPLAIPTPSLTYSLADSLANASCQIIPPLLVQPMQ
FSNSSCLSSPFINDTEQIDLGAVTFTNCTSVANVSSPLCALNGSVFLCGNNMAYTYLPQ
NWTGLCVQASLLPDIDIIPGDEPVPIPAIDHYIHRPKRAVQFIPLLAGLGITAAFTTGA
TGLGVSVTQYTKLSHQLISDVQVLSGTIQDLQDQVDSLAEVVLQNRRGLDLLTAEQGGI
CLALQEKCCFYANKSGIVRNKIRTLQEELQKRRESLASNPLWTGLQGFLPYLLPLLGPL
LTLLLILTIGPCVF"
CDS 2813..2914
/codon_start=1
/label=ENV from the MLV 4070A
/note="cytoplasmic tail domain derived from the MLV 4070A"
/translation="NRLVQFVKDRISVVQALVLTQQYHQLKPIEYEP"
polyA_signal 3044..3165
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(3172..3627)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3654..3758
/label=AmpR promoter
promoter 3760..4117
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4152..4943
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5134..5153)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5178..5225
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5554..6142
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"