pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE vector (V013750)

Basic Vector Information

Vector Name:
pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE
Antibiotic Resistance:
Ampicillin
Length:
11866 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Puro
Promoter:
EF-1α
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE vector Vector Map

pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE11866 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000115003' LTRHIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSEF-1-alpha promoterluciferaseIRES2EGFPPGK promoterPuroRWPRE5' LTRlac promoterCAP binding siteoriAmpRAmpR promoterSV40 poly(A) signal

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 ℃.

pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_17150       11866 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11866)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2027..2144
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2338..3519
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3535..5184
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    5214..5788
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             5818..6534
                     /label=EGFP
                     /note="enhanced GFP"
     promoter        6567..7065
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             7125..7721
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    7787..8375
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8582..9215
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(9392..9422)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9437..9458)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9746..10334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10508..11365)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(11366..11470)
                     /label=AmpR promoter
     polyA_signal    11518..11652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"

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