pLenti-DsRed_IRES_SNCA:EGFP vector (V017172)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017172 pLenti-DsRed_IRES_SNCA:EGFP 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:
pLenti-DsRed_IRES_SNCA:EGFP
Antibiotic Resistance:
Ampicillin
Length:
10354 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Neo/G418
Promoter:
CMV
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pLenti-DsRed_IRES_SNCA:EGFP vector Map

pLenti-DsRed_IRES_SNCA:EGFP10354 bp500100015002000250030003500400045005000550060006500700075008000850090009500100005' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSCMV enhancerCMV promoterDsRed-ExpressIRES2EGFPT7 promoterPGK promoterNeoR/KanRWPRE3' LTR (Delta-U3)AmpR promoterAmpRori3' LTR

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.

pLenti-DsRed_IRES_SNCA:EGFP vector Sequence

LOCUS       V017172                10354 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017172
VERSION     V017172
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10354)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             376..633
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    680..805
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1302..1535
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1720..1764
                     /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             1913..1954
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2057..2174
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2226..2529
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2530..2733
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2843..3517
                     /label="DsRed-Express"
                     /note="rapidly maturing tetrameric variant of DsRed
                     fluorescent protein (Bevis and Glick, 2002)"
     misc_feature    3546..4130
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4571..5287
                     /label="EGFP"
                     /note="enhanced GFP"
     promoter        complement(5311..5329)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5339..5814
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             5835..6626
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    6743..7331
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7406..7639
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        7844..7948
                     /label="AmpR promoter"
     CDS             7949..8806
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8980..9568
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    9979..10000
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     LTR             10000..10354
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"