pLVX-PAmCherry-N1 vector (V012716)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V012716	pLVX-PAmCherry-N1	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

The pLVX-PAmCherry-N1 is a Lentiviral Gene Expression with Photoactivatable Fluorescence. The pLVX-PAmCherry-C1 vector is an HIV-1-based lentiviral expression vector designed to efficiently infect cells and express your gene of interest. The lentiviral particles derived from this vector encode a photoactivatable fluorescent protein, PAmCherry, fused to your gene of interest. PAmCherry, a mutant of mCherry, exhibits no fluorescence until photoactivated by exposure to light within the 350-400 nm wavelength range. Upon activation, PAmCherry emits light at 564 nm (excitation) and 595 nm (emission). Genes cloned into the vector's MCS will be expressed as fusion proteins at the C-terminus of PAmCherry, provided they are in-frame and lack intervening stop codons. This allows for precise expression control and visualization of your protein of interest.

Vector Name:: pLVX-PAmCherry-N1

Antibiotic Resistance:: Ampicillin

Length:: 8777 bp

Type:: Viral Expression & Packaging Vectors

Replication origin:: ori

Source/Author:: Clontech

Selection Marker:: Puromycin

Copy Number:: High copy number

Promoter:: mPGK

Cloning Method:: Enzyme Cut

Fusion Tag:: PAmCherry

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

Expression Method:: Constiutive, Stable

pLVX-PAmCherry-N1 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.

References

Wang X, Wu Z, Li Y, Yang Y, Xiao C, Liu X, Xiang X, Wei J, Shao D, Liu K, Deng X, Wu J, Qiu Y, Li B, Ma Z. p53 promotes ZDHHC1-mediated IFITM3 palmitoylation to inhibit Japanese encephalitis virus replication. PLoS Pathog. 2020 Oct 27;16(10):e1009035. doi: 10.1371/journal.ppat.1009035. PMID: 33108395; PMCID: PMC7647115.

pLVX-PAmCherry-N1 vector Sequence

LOCUS       V012716                 8777 bp    DNA     circular SYN 08-APR-2021
DEFINITION  Exported.
ACCESSION   V012716
VERSION     V012716
KEYWORDS    pLVX-PAmCherry-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8777)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8777)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8777
                     /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"
                     /label="Protein Tat"
     misc_feature    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2201..2504
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    2802..2867
                     /label="MCS"
                     /note="multiple cloning site"
     CDS             2874..3581
                     /label="PAmCherry1"
                     /note="photoactivatable monomeric derivative of DsRed
                     fluorescent protein (Subach et al., 2009)"
     promoter        3612..4111
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4132..4728
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4745..5333
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5540..6173
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6302..6318)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6326..6342)
                     /label="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(6350..6380)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6395..6416)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6704..7292)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7466..8323)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8324..8428)
                     /label="AmpR promoter"
     polyA_signal    8476..8610
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"