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pShuttle-IRES-hrGFP2 vector (V013058)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013058 pShuttle-IRES-hrGFP2 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:
pShuttle-IRES-hrGFP2
Antibiotic Resistance:
Kanamycin
Length:
9023 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Adenovirus
Promoter:
CMV
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pShuttle-IRES-hrGFP2 vector Map

Copy Sequence View Map
pShuttle-IRES-hrGFP29023 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800NeoR/KanRM13 oriITRAd5 PsiCMV enhancerCMV promoter3xHAIRES2hrGFPSV40 poly(A) signalHexon-interlacing proteinPackaging protein 1Early 4 ORF1 proteinITRori

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.

pShuttle-IRES-hrGFP2 vector Sequence

LOCUS       V013058                 9023 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013058
VERSION     V013058
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9023)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             69..860
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(911..1291)
                     /direction=LEFT
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     repeat_region   1329..1431
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     misc_signal     1519..1669
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        1727..2030
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2031..2234
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2326..2406
                     /label="3xHA"
                     /note="three tandem HA epitope tags"
     misc_feature    2442..3025
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             3026..3742
                     /label="hrGFP"
                     /note="humanized Renilla green fluorescent protein"
     polyA_signal    3887..4008
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             4102..4521
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(4587..5921)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     CDS             complement(6506..6889)
                     /note="Early 4 ORF1 protein from Human adenovirus C
                     serotype 2. Accession#: P03242"
                     /label="Early 4 ORF1 protein"
     repeat_region   7199..7301
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     rep_origin      complement(7561..8149)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"