Basic Vector Information
- Vector Name:
- L202 pPLVPTH2-tTR-KRAB-Cerulean-scrambled-shRNA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12060 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
L202 pPLVPTH2-tTR-KRAB-Cerulean-scrambled-shRNA 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 ℃.
L202 pPLVPTH2-tTR-KRAB-Cerulean-scrambled-shRNA vector Sequence
LOCUS 62056_1385 12060 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 12060) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..12060 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1299..1532 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1716..1760 /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 1909..1950 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 2027..2144 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2183..2682 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 2745..3461 /label=mCerulean /note="enhanced monomeric variant of CFP (Rizzo et al., 2004)" protein_bind complement(3466..3490) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" misc_feature 3511..4073 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4078..4698 /label=TetR /note="tetracycline repressor TetR" CDS 4699..4719 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" CDS 4822..5016 /label=KRAB /note="Kruppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)" misc_feature 5192..5780 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" protein_bind 5919..6189 /label=tetracycline response element /note="contains seven copies of the tetracycline operator tetO" promoter 6380..6592 /label=H1 promoter /note="human H1 RNA promoter" LTR 6674..6854 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(6879..6897) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" promoter 7685..7789 /label=AmpR promoter CDS 7790..8647 /label=AmpR /note="beta-lactamase" rep_origin 8821..9409 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 9655..9984 /label=SV40 promoter /note="SV40 enhancer and early promoter" intron 11118..11183 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 11313..11333 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" polyA_signal 11758..11892 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal"
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