pLV-EF1a-IgκL-HA-OKT3scFv(opt-human)-Myc-PDGFRβ-Puro vector (V014575)

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Cat No.	Plasmid Name	Availability	Add to cart
V014575	pLV-EF1a-IgκL-HA-OKT3scFv(opt-human)-Myc-PDGFRβ-Puro	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: pLV-EF1a-IgκL-HA-OKT3scFv(opt-human)-Myc-PDGFRβ-Puro

Antibiotic Resistance:: Ampicillin

Length:: 8569 bp

Type:: Gene expression

Replication origin:: ori

Host:: Mammalian cells, Lentivirus

Selection Marker:: Puro

Promoter:: EF-1α

Growth Strain(s):: Stbl3

Growth Temperature:: 30℃

pLV-EF1a-IgκL-HA-OKT3scFv(opt-human)-Myc-PDGFRβ-Puro vector Map

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Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pLV-EF1a-IgκL-HA-OKT3scFv(opt-human)-Myc-PDGFRβ-Puro vector Sequence

LOCUS       V014575                 8569 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014575
VERSION     V014575
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8569)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             117..750
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    797..922
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1415..1648
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1833..1877
                     /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             2026..2067
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2164..2281
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2409..3587
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     sig_peptide     3613..3675
                     /label="Ig-kappa leader"
                     /note="leader sequence from mouse immunoglobulin kappa
                     light chain"
     CDS             3676..3702
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             4483..4629
                     /label="PDGFR-beta TM domain"
                     /note="transmembrane domain from platelet derived growth
                     factor receptor beta"
     misc_feature    4685..5271
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             5291..5887
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             6093..6326
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6483..6707
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        6803..6907
                     /label="AmpR promoter"
     CDS             6908..7765
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7939..8527
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"