pLVX-EF1α-IRES-mCherry

$216.00

  • Model: PVT2309
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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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