Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001709 | EJ2GFP-puro | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
This is a mammalian expression vector, with EJ2GFP egfp-based chromosomal break reporter and puromycin selection marker.
EJ2GFP-puro vector Vector Map
Plasmid Resuspension 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 ℃.
References
- Bennardo N, Cheng A, Huang N, Stark JM. Alternative-NHEJ is a mechanistically distinct pathway of mammalian chromosome break repair. PLoS Genet. 2008 Jun 27;4(6):e1000110.
EJ2GFP-puro vector Sequence
LOCUS Exported 7642 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS EJ2GFP-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7642)
AUTHORS Bennardo N, Cheng A, Huang N, Stark JM
TITLE Alternative-NHEJ is a mechanistically distinct pathway of mammalian
chromosome break repair.
JOURNAL PLoS Genet. 2008 Jun 27;4(6):e1000110. doi:
10.1371/journal.pgen.1000110.
PUBMED 18584027
REFERENCE 2 (bases 1 to 7642)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS
Genet."; date: "2008-06-27"; volume: "4(6):e1000110. doi"; pages: "
10.1371/journal.pgen.1000110"
FEATURES Location/Qualifiers
source 1..7642
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 658..1674
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1598..1620
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1682..1701
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1763..1783
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1805..1825
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 2074..2083
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2080..2799
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 2080..2796
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2125..2146)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2386..2405)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2733..2754
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3004..3023)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3069..3124
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3123..3142)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
promoter complement(3482..3500)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3483..3500)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3581..3600)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3700..3722
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3760..3778)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3846..3950
/gene="bla"
/label=AmpR promoter
CDS 3951..4811
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4169..4188)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4982..5570
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5471..5490
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5724..5741
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 5828..5847
/label=T7
/note="T7 promoter, forward primer"
promoter 5828..5846
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 5884..6381
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(5919..5942)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 6314..6333
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 6400..6999
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(6400..6419)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6896..6916
/label=Puro-F
/note="Puromycin resistance gene, forward primer"