EJ2GFP-puro vector (V001709)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001709 EJ2GFP-puro In stock (lyophilized plasmid)

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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.

      • Vector Name:
      • EJ2GFP-puro
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7642 bp
      • Type:
      • Mammalian Expression
      • Replication origin:
      • ori
      • Selection Marker:
      • Puromycin
      • Promoter:
      • CAG
      • 5' Primer:
      • ttcctacagctcctgggcaacg
      • 3' Primer:
      • ttttggcagagggaaaaaga
      • Growth Strain(s):
      • Stbl3
      • Growth Temperature:
      • 37℃

EJ2GFP-puro vector Vector Map

EJ2GFP-puro7642 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500CMV enhancerchicken beta-actin promoterchimeric intronpCAG-FSV40 NLSSV40 NLSEGFPBglob-pA-Rbeta-globin poly(A) signalSP6 promoterpRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRoriL4440T7 promoterPGK promoterPuroRCAG

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_765        7642 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7642)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS
            Genet."; date: "2008-06-27"; pages: "
            10.1371/journal.pgen.1000110"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..657
                     /label=chicken beta-actin promoter
     intron          658..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     CDS             1763..1783
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1805..1825
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             2080..2796
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     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(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
                     /label=AmpR promoter
     CDS             3951..4808
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4982..5570
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5724..5741
                     /label=L4440
                     /note="L4440 vector, 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"
     CDS             6400..6996
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        7640..7642
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin promoter"