pLV-UBC-HBx(HBV)-EGFP vector (V014723)

Price Information

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V014723 pLV-UBC-HBx(HBV)-EGFP In stock, instant shipping

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

Basic Vector Information

Vector Name:
pLV-UBC-HBx(HBV)-EGFP
Antibiotic Resistance:
Ampicillin
Length:
10402 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Zeo
Promoter:
UbC
Growth Strain(s):
stbl3
Growth Temperature:
37℃

pLV-UBC-HBx(HBV)-EGFP vector Map

pLV-UBC-HBx(HBV)-EGFP10402 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000BleoRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoterCMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSUbC promoterEGFPWPREbGH poly(A) signalf1 oriSV40 promoterEM7 promoter

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-UBC-HBx(HBV)-EGFP vector Sequence

LOCUS       V014723                10402 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014723
VERSION     V014723
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10402)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    417..550
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(635..665)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(680..701)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(989..1577)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1751..2608)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(2609..2713)
                     /label="AmpR promoter"
     enhancer        2979..3358
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3359..3561
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             3576..3756
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3803..3928
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    4421..4654
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             4839..4883
                     /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             5032..5073
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    5181..5298
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        5363..6574
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             7098..7814
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    7844..8432
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    9160..9384
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9430..9858
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        10005..10201
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     promoter        10249..10296
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             join(10315..10402,1..284)
                     /label="BleoR"
                     /note="antibiotic-binding protein"