pLV3-CMV-mCherry-Linker-PKP1 vector (V014125)

Price Information

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V014125 pLV3-CMV-mCherry-Linker-PKP1 In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pLV3-CMV-mCherry-Linker-PKP1
Antibiotic Resistance:
Ampicillin
Length:
10357 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Puro
Promoter:
RSV
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pLV3-CMV-mCherry-Linker-PKP1 vector Vector Map

pLV3-CMV-mCherry-Linker-PKP110357 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000RSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSCMV promotermCherryPlakophilin-1EF-1-alpha core promoter5' LTR (truncated)PuroRWPRE3' LTR (Delta-U3)SV40 poly(A) signalSV40 orilac promoterCAP binding siteoriAmpRAmpR promoterM13 fwd

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.

pLV3-CMV-mCherry-Linker-PKP1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V014125                10357 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014125
VERSION     V014125
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10357)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /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             1686..1727
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    1804..1920
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2010..2213
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2301..3008
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             3042..5282
                     /gene="PKP1"
                     /label="Plakophilin-1"
                     /note="Plakophilin-1 from Homo sapiens. Accession#: Q13835"
     promoter        5327..5538
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             5551..5819
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             5846..6442
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    6452..7040
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7114..7347
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7419..7553
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7559..7694
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(7780..7810)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7825..7846)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8134..8722)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8896..9753)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9754..9858)
                     /label="AmpR promoter"
     primer_bind     10332..10348
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"