Basic Vector Information
- Vector Name:
- pLenti CMV/TO GFP-Zeo DEST (719-1)
- Antibiotic Resistance:
- Ampicillin, Chloramphenicol
- Length:
- 10150 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Zeo
- Promoter:
- RSV
- Growth Strain(s):
- DB3.1
- Growth Temperature:
- 37℃
pLenti CMV/TO GFP-Zeo DEST (719-1) 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 GFP-Zeo DEST (719-1) vector Sequence
LOCUS 62056_13820 10150 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10150) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..10150 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 26..130 /label=AmpR promoter CDS 131..988 /label=AmpR /note="beta-lactamase" rep_origin 1162..1750 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2038..2059 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2074..2104 /label=lac promoter /note="promoter for the E. coli lac operon" promoter 2173..2191 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2219..2445 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" misc_feature 2673..2798 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3291..3524 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3709..3753 /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 3902..3943 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 4020..4136 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 4463..4662 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 4667..4685 /label=tet operator /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4747..4871 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 4896..4926 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 4980..5636 /label=CmR /note="chloramphenicol acetyltransferase" CDS 5981..6283 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" protein_bind complement(6327..6451) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" misc_feature 6482..7070 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" enhancer 7145..7448 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7449..7652 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 7697..8413 /label=EGFP /note="enhanced GFP" LTR 8887..9120 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 9192..9326 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 9353..9488 /label=SV40 ori /note="SV40 origin of replication" promoter complement(9509..9527) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase"
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