pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro vector (V015004)

Basic Vector Information

Vector Name:
pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro
Antibiotic Resistance:
Ampicillin
Length:
10471 bp
Type:
Protein expression, Tetracycline inducible
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Puro
Promoter:
hPGK
Growth Temperature:
37℃

pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro vector Vector Map

pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro10471 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000SV40 poly(A) signalAmpR promoterAmpRoriCAP binding sitelac promoter5' LTRWPREbGH poly(A) signalPuroRIRES2Tet-On(R) 3GhPGK promoterTRE3GS promoterEGFPSV40 poly(A) signalcPPT/CTSProtein Tatgp41 peptideRREHIV-1 Psi3' LTR

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V015004                10471 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015004
VERSION     V015004
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10471)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(216..350)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        398..502
                     /label="AmpR promoter"
     CDS             503..1360
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1534..2119
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2407..2428
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2443..2473
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     LTR             complement(2650..3283)
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     misc_feature    complement(3490..4078)
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(4092..4316)
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4344..4940)
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    complement(4960..5546)
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             complement(5562..6305)
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(6324..6834)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        6851..7218
                     /label="TRE3GS promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
     CDS             7257..7973
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    complement(8151..8285)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(8328..8445)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(8517..8558)
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     CDS             complement(8707..8751)
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     misc_feature    complement(8936..9169)
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    complement(9666..9791)
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(9838..10471)
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"

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