pLVX-EF1α-IRES-puro PVT2308 2ug
pLVX-EF1α-IRES-puro Information
Promoter: EF-1 alpha
Replicon: pUC ori
Terminator: SV40 poly (A) signal
Plasmid classification: virus series, lentivirus cloning vector
Plasmid size: 8825bp
Prokaryotic resistance: Amp
Selection marker: Puro
Clone strain: Stbl3
Culture conditions: 37 C, aerobic LB
Expression host: lactation cells
Induction mode: no need to induce, transient expression.
Primers for 5'sequencing: primers designed based on sequences
Primers for 3'sequencing: primers designed based on sequences
pLVX-EF1α-IRES-puro Description
pLVX-EF1α-IRES-Puro contains all of the viral processing elements necessary for the production of replicationincompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA, leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus. Finally, pLVX-EF1α-IRES-Puro also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import ofthe viral genome, resulting in improved vector integration and more efficient transduction. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.
pLVX-EF1α-IRES-puro Multiple cloning site
pLVX-EF1α-IRES-puro Sequence
LOCUS Exported 8825 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 22
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8825)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..8825
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 2338..3519
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2568..3510
/note="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
misc_feature 3575..4148
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4181..4780
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4794..5382
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5265..5276)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5589..6222
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6350..6366)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6374..6390
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
promoter complement(6398..6428)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6443..6464
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6752..7340)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7511..8371)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
温馨提示：不可用于临床ZL。
