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pIRES vector (V011109)

Price Information

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V011109 pIRES 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:
pIRES
Antibiotic Resistance:
Ampicillin
Length:
6105 bp
Type:
Mammalian Expression Vectors
Replication origin:
ori
Source/Author:
Clontech
Selection Marker:
Neomycin/G418(Geneticin)
Copy Number:
High copy number
Promoter:
CMV
Growth Strain(s):
DH10B

pIRES vector Map

Copy Sequence View Map
pIRES6105 bp30060090012001500180021002400270030003300360039004200450048005100540057006000CMV enhancerCMV promoterchimeric intronMCS AIRES2T3 promoterSV40 poly(A) signalf1 oriSV40 promoterNeoR/KanRpoly(A) signalAmpR 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.

pIRES vector Sequence

LOCUS       pIRES.        6105 bp DNA     circular SYN 01-JAN-1980
DEFINITION  IRES-containing vector for expressing two genes in mammalian cells 
            from the same bicistronic transcript.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6105)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6105)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1085..1107
                     /label=MCS A
                     /note="MCS A"
                     /note="multiple cloning site"
     misc_feature    1152..1738
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1737..1770
                     /label=MCS B
                     /note="MCS B"
                     /note="multiple cloning site"
     promoter        complement(1774..1791)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase
                     (shorter by one base than the standard T3 promoter)"
     polyA_signal    complement(1809..1930)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2116..2571
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2688..3045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3096..3887
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3954..4002
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        4308..4412
                     /label=AmpR promoter
     CDS             4413..5270
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5444..6032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"