Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V012797 | pNL4-3.Luc.R-E- | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
Firefly luciferase gene was inserted into the pNL4-3 nef gene. Two frameshifts (5'Env and Vpr aa 26) render this clone Env- and Vpr-. Competent for a single round of replication. Requires cotransfection with env expression vector to produce infectious virus. Virus can be produced by transfecting 2x10^6 293 or 293T cells with 20 ug of the NL4-3 DNA, or with 10 ug NL4-3 DNA and 10 ug env expression vector DNA. Transfections can be performed in a 10 cm^2 tissue culture dish using standard calcium phosphate protocols. Virus is typically harvested 48 hours post-transfection. Infections should be performed in a total volume of 0.5 ml. Amphotropic pseudotypes generally have much higher infectivity than those bearing HIV-1 env. Cultures infected with the luciferase viruses can be lysed 2-5 days post-infection and assayed using commercial lysis buffer and luciferase reagents
Basic Vector Information | |||
---|---|---|---|
Vector Name | pNL4-3.Luc.R-E- | Antibiotic Resistance | Ampicillin |
Length | 16390 bp | Type | Viral Expression & Packaging Vectors |
Source | Dr. Nathaniel Landau, Aaron Diamond AIDS Research Center, The Rockefeller University. | Copy Number | High copy number |
Expression Method | Constiutive, Stable |
pNL4-3.Luc.R-E- vector Vector Map
Plasmid Resuspension 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 ℃.
pNL4-3.Luc.R-E- vector Sequence
LOCUS Exported 16390 bp ds-DNA circular SYN 08-OCT-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pnl4-3-luc-r-e- SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16390) AUTHORS Triple Threat TITLE Direct Submission FEATURES Location/Qualifiers source 1..16390 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 669..773 /gene="bla" /note="AmpR promoter" CDS 774..1634 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1805..2393 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" LTR 4204..4837 /note="3' LTR" /note="3' long terminal repeat (LTR) from HIV-1" CDS complement(5027..6673) /codon_start=1 /gene="luc" /product="firefly luciferase" /note="luciferase" /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH IEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA NDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF VDEVPKGLTGKLDARKIREILIKAKKGGKSKL" CDS complement(6937..6963) /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /note="NES" /translation="LPPLERLTL" misc_feature 7486..7719 /note="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS complement(10382..13393) /codon_start=1 /gene="pol" /product="pol protein from human immunodeficiency virus 1" /note="HIV-1 pol" /translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG GIGGFIKVRQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD STKWRKLVDFRELNKRTQDFWEVQLGIPHPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH KGIGGNEQVDKLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ MAGDDCVASRQDED" misc_feature 10580..10697 /note="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(13186..14688) /codon_start=1 /gene="gag" /product="gag protein from human immunodeficiency virus 1" /note="HIV-1 gag" /translation="MGARASVLSGGELDKWEKIRLRPGGKKQYKLKHIVWASRELERFA VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTIAVLYCVHQRIDVKDTKEALDKI EEEQNKSKKKAQQAAADTGNNSQVSQNYPIVQNLQGQMVHQAISPRTLNAWVKVVEEKA FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRLHPVHAGP IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ" LTR 14844..15101 /note="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1"