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pLVX-EF1α-DsRed-Monomer-N1 vector (V012728#)

Basic Vector Information

pLVX-EF1α-DsRed-Monomer-N1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the N-terminus of the fluorescent protein DsRed-Monomer. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells), without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing DsRed-Monomer fusions, without time-consuming drug and clonal selection.

Basic Vector Information
Vector Name pLVX-EF1α-DsRed-Monomer-N1 Antibiotic Resistance Ampicillin
Length 9843 bp Type Viral Expression & Packaging Vectors
Source Clontech Selection Marker Puromycin
Copy Number High copy number Promoter EF1α/ EF1a
Cloning Method Enzyme Cut Fusion Tag DsRed-Monomer
Expression Method Constiutive, Stable

pLVX-EF1α-DsRed-Monomer-N1 vector Vector Map

pLVX-EF1α-DsRed-Monomer-N1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9483 bp ds-DNA     circular SYN 08-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-EF1-alpha-DsRed-Monomer-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9483)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9483
                     /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    3525..3605
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             3613..4290
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /note="DsRed-Monomer"
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     promoter        4319..4818
                     /note="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4839..5438
                     /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    5452..6040
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5923..5934)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     LTR             6247..6880
                     /note="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7008..7024)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7032..7048
                     /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(7056..7086)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7101..7122
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7410..7998)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8169..9029)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9030..9134)
                     /gene="bla"
                     /note="AmpR promoter"
     polyA_signal    9182..9316
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"

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