Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010386 | pRetroX-IRES-ZsGreen1 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pRetroX-IRES-ZsGreen1 is a bicistronic, fl uorescent, retroviral vector that allows both a gene of interest and the ZsGreen1 gene to be translated from a single bicistronic mRNA. pRetroXIRES-ZsGreen1 is designed for effi cient delivery and selection (by fl ow cytometry or other methods) of stably transduced mammalian cells expressing the ZsGreen1 fl uorescent protein and the protein of interest. This vector can be used to obtain stable cell lines without timeconsuming drug and clonal selection. ZsGreen1 is derived from the reef coral Zoanthus sp. green fl uorescent protein (ZsGreen) and is easily detected with standard FITC fi lter sets.
- Vector Name:
- pRetroX-IRES-ZsGreen1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5953 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- CMV
pRetroX-IRES-ZsGreen1 vector Map
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.
pRetroX-IRES-ZsGreen1 vector Sequence
LOCUS pRetroX-IRES-ZsG 5953 bp DNA circular SYN 01-JAN-1980
DEFINITION Bicistronic retroviral vector for expressing a gene together with
the fluorescent protein ZsGreen1.
ACCESSION .
VERSION .
KEYWORDS pRetroX-IRES-ZsGreen1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5953)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5953)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5953
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..592
/label=5' LTR
/note="5' LTR"
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 655..1012
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
intron 1075..1278
/label=EF-1-alpha intron A
/note="truncated intron upstream of the start codon of
human EF-1-alpha"
exon 1279..1308
/label=EF-1-alpha exon
/note="truncated exon 2 from human EF-1-alpha, beginning
with a GT-to-CC mutation that reduces splicing efficiency"
misc_feature 1362..1389
/label=MCS
/note="MCS"
/note="multiple cloning site"
misc_feature 1393..1972
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 1967..2659
/label=ZsGreen1
/note="Zoanthus green fluorescent protein"
LTR 2716..3309
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(3325..3341)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3349..3365)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3373..3403)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3418..3439)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3727..4315)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4489..5346)
/label=AmpR
/note="beta-lactamase"
promoter complement(5347..5451)
/label=AmpR promoter
primer_bind 5925..5941
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"