Basic Vector Information
pLenti CMV/TO eGFP Puro (w159-1) 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 ℃.
pLenti CMV/TO eGFP Puro (w159-1) vector Sequence
LOCUS pLenti_CMV/TO_eG 8771 bp DNA circular SYN 13-MAY-2021 DEFINITION 3rd gen lentiviral eGFP expression vector, CMV/TO promoter, Puro . ACCESSION . VERSION . KEYWORDS pLenti CMV/TO eGFP Puro (w159-1). SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8771) AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. TITLE A versatile viral system for expression and depletion of proteins in mammalian cells. JOURNAL PLoS One. 2009 Aug 6;4(8):e6529. PUBMED 19657394 REFERENCE 2 (bases 1 to 8771) TITLE Direct Submission REFERENCE 3 (bases 1 to 8771) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One."; date: "2009-08-6"; volume: "4(8)"; pages: "e6529" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..8771 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 27..131 /label=AmpR promoter CDS 132..989 /label=AmpR /note="beta-lactamase" rep_origin 1163..1751 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1905..1922 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2039..2060 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2075..2105 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2113..2129 /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2137..2153 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2174..2192 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2220..2447 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 2448..2628 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2672..2797 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3290..3523 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3707..3751 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" CDS 3900..3941 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 4012..4129 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 4152..4455 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4456..4659 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4656..4680 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 4764..4788 /label=attB1 /note="recombination site for the Gateway(R) BP reaction" CDS 4854..5570 /label=EGFP /note="enhanced GFP" protein_bind complement(5598..5622) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" misc_feature 5653..6241 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" promoter 6255..6754 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 6775..7371 /label=PuroR /note="puromycin N-acetyltransferase" LTR 7512..7745 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7817..7951 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 7957..8092 /label=SV40 ori /note="SV40 origin of replication" promoter complement(8125..8143) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8153..8169) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 8314..8769 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
This page is informational only.