pLVX-EF1α-IRES-mCherry
PVT2309 2ug
pLVX-EF1α-IRES-mCherry Informaiton
Promoter: EF-1 alpha promoter
Replicon: pUC ori
Terminator: SV40 poly (A) signal
Plasmid classification: virus series, lentivirus cloning vector
Prokaryotic resistance: ampicillin Amp
Selection marker: red fluorescence
Clone strain: Escherichia coli Stbl3
Culture conditions: 37 C, aerobic, LB
Expression host: lactation cells
Induction mode: no need to induce, transient expression.
5'sequencing primers: WPRE-R:CATAGCGTAAAAGGAGCAACA
Primers for 3'sequencing: primers designed based on sequences
Remarks: there is a difference from the sequence of Clontech.
pLVX-EF1α-IRES-mCherry Description
Plvx-ef1 alpha-IRES mCherry is a bigenic lentiviral expression vector that can be used to transduce mammalian cells to produce high titers of lentiviruses, including primary and stem cells, in almost any division or non-division. The vector contains an internal ribosome entry site (IRES) that allows an interesting gene and red fluorescent protein mCherry to be simultaneously transcribed from a single mRNA expression. Transcriptionally expressed human elongation factor-1 alpha (EF-1 alpha) promoters are driven and continue to be activated even when the genome of the host cell is stably integrated into the vector. Stable transcription allows the monitoring of various cellular processes, such as primary or stem cell differentiation, not to be silenced with CMV promoters. In addition, vectors allow efficient flow cytometry to detect the expressed proteins, mCherry, and benefits of stable or transient transfection of mammalian cells without time-consuming drug and cloning options.
pLVX-EF1α-IRES-mCherry Sequence
LOCUS Exported 8935 bp ds-DNA circular SYN 07-SEP-2016
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pLVX-EF1-alpha-IRES-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8935)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, September 7, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..8935
/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..4147
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4180..4890
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/note="mCherry"
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
misc_feature 4904..5492
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5375..5386)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5699..6332
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6460..6476)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6484..6500
/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(6508..6538)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6553..6574
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6862..7450)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7621..8481)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
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