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pHR_Gal4UAS_EGFP_PGK_mCherry vector (V014761)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014761 pHR_Gal4UAS_EGFP_PGK_mCherry 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:
pHR_Gal4UAS_EGFP_PGK_mCherry
Antibiotic Resistance:
Ampicillin
Length:
10783 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Promoter:
mPGK
Growth Temperature:
37℃

pHR_Gal4UAS_EGFP_PGK_mCherry vector Map

Copy Sequence View Map
pHR_Gal4UAS_EGFP_PGK_mCherry10783 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500small t intronSV40 NLSSV40 poly(A) signal3' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSEGFPPGK promotermCherryWPRE3' LTR (Delta-U3)SP6 promoterAmpR promoterAmpRori

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.

pHR_Gal4UAS_EGFP_PGK_mCherry vector Sequence

LOCUS       V014761                10783 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014761
VERSION     V014761
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10783)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          455..520
                     /label="small t intron"
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             650..670
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    1095..1229
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             1397..2030
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2077..2202
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    2699..2932
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3117..3161
                     /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             3310..3351
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    3446..3563
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             3894..4610
                     /label="EGFP"
                     /note="enhanced GFP"
     promoter        4736..5235
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             5256..5963
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    6043..6631
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6741..6974
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(6999..7017)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7805..7909
                     /label="AmpR promoter"
     CDS             7910..8767
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8941..9529
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"