Basic Vector Information
- Vector Name:
- L201 pLVPTH2-tTR-KRAB-Cerulean-scrambled-shRNA-Control
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12031 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
L201 pLVPTH2-tTR-KRAB-Cerulean-scrambled-shRNA-Control 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 ℃.
L201 pLVPTH2-tTR-KRAB-Cerulean-scrambled-shRNA-Control vector Sequence
LOCUS 62056_1380 12031 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 12031)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12031
/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 6645..6825
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(6850..6868)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 7656..7760
/label=AmpR promoter
CDS 7761..8618
/label=AmpR
/note="beta-lactamase"
rep_origin 8792..9380
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 9626..9955
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
intron 11089..11154
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 11284..11304
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 11729..11863
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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