pGIPZ vector (V010484)

Price Information

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V010484 pGIPZ 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:
pGIPZ
Antibiotic Resistance:
Ampicillin
Length:
11688 bp
Type:
Viral Expression & Packaging Vectors
Replication origin:
ori
Source/Author:
Open Biosystems
Copy Number:
High copy number
Promoter:
SV40
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pGIPZ vector Map

pGIPZ11688 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000115003' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSBleoREM7 promoterCMV enhancerCMV promoterTurboGFPPuroR5' miR-30a3' miR-30aT7 promoterWPRE3' LTR (Delta-U3)bGH poly(A) signalf1 oriSV40 promoterHygRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoterSV40 poly(A) signal

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.

pGIPZ vector Sequence

LOCUS       V010484                11688 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010484
VERSION     V010484
KEYWORDS    pGIPZ
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11688)
  AUTHORS   Open Biosystems
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 11688)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11688
                     /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    2063..2180
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(2248..2619)
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     promoter        complement(2639..2686)
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     enhancer        2791..3094
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3095..3298
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             3390..4085
                     /label="TurboGFP"
                     /note="improved green fluorescent protein from Pontellina
                     plumata (Evdokimov et al., 2006)"
     misc_feature    4115..4697
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4696..5289
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     ncRNA           5299..5392
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     ncRNA           5418..5530
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     promoter        complement(5625..5643)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5663..6251
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6458..6691
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6798..7022
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7068..7496
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7510..7839
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             7888..8910
                     /label="HygR"
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    9043..9176
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(9261..9291)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9306..9327)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9615..10203)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10377..11234)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(11235..11339)
                     /label="AmpR promoter"
     polyA_signal    11387..11521
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"