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pLVX-EF1α-AcGFP1-C1 vector (V012730#)

Basic Vector Information

pLVX-EF1α-AcGFP1-C1 is an HIV-1-based, lentiviral expression vector designed to constitutively express a protein of interest fused to the C-terminus of AcGFP1, a green fluorescent protein derived from Aequorea coerulescens. The excitation and emission maxima of native AcGFP1 are 475 nm and 505 nm, respectively. Stable, constitutive expression of the fusion protein is driven by the EF1α promoter (PEF1α), which continues to be constitutively active even after the vector integrates into the host cell genome (1). pLVX-EF1α-AcGFP1-C1 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 (2), 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 (3). Finally, pLVX-EF1α-AcGFP1-C1 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 of the viral genome, resulting in improved vector integration and more efficient transduction (4). In addition to lentiviral elements, pLVX-EF1α-AcGFP-C1 contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.

Basic Vector Information
Vector Name pLVX-EF1α-AcGFP1-C1 Antibiotic Resistance Ampicillin
Length 9519 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 AcGFP1
Expression Method Constiutive, Stable

pLVX-EF1α-AcGFP1-C1 vector Vector Map

pLVX-EF1α-AcGFP1-C1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9519 bp ds-DNA     circular SYN 08-APR-2021
DEFINITION  synthetic circular DNA
KEYWORDS    pLVX-EF1-alpha-AcGFP1-C1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9519)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9519
                     /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="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
     misc_feature    2028..2143
                     /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 
     CDS             3539..4255
                     /product="Aequorea coerulescens GFP"
                     /note="mammalian codon-optimized"
     misc_feature    4256..4321
                     /note="multiple cloning site"
     promoter        4354..4853
                     /note="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4874..5473
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="confers resistance to puromycin"
     misc_feature    5487..6075
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5958..5969)
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
     LTR             6282..6915
                     /note="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7044..7060)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
     protein_bind    7068..7084
                     /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(7092..7122)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7137..7158
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7446..8034)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(8205..9065)
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(9066..9170)
                     /note="AmpR promoter"
     polyA_signal    9218..9352
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"

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