MS2-P65-HSF1_GFP vector (V012320)

Basic Vector Information

      • Vector Name:
      • MS2-P65-HSF1_GFP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 11409 bp
      • Type:
      • CRISPR Plasmids
      • Source/Author:
      • Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
      • Copy Number:
      • High copy number

MS2-P65-HSF1_GFP vector Vector Map

MS2-P65-HSF1_GFP11409 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000CMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSEF-1-alpha promoterMS2/P65/HSF1T2AEGFPWPRE5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

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 ℃.

MS2-P65-HSF1_GFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       MS2-P65-HSF1_GFP       11409 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Zhang lab vector for expressing GFP together with an MS2 RNA-binding
            domain fused to the p65 and HSF1 transcription factors.
ACCESSION   .
VERSION     .
KEYWORDS    MS2-P65-HSF1_GFP.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11409)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F.
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature 2015;517:583-8.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 11409)
  AUTHORS   Zhang Lab / Addgene #61423
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 11409)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature"; 
            date: "2015"; volume: "517"; pages: "583-8"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             2291..2332
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2682..3859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3872..5290
                     /label=MS2/P65/HSF1
                     /note="Fusion protein of MS2 bacteriophage coat protein, 
                     NF-kappaB trans-activating subunit p65 and human heat-shock
                     factor 1 activation domain"
     CDS             5306..5359
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=2A peptide from Thosea asigna virus capsid
                     protein
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5360..6073
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    6101..6689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7214..7394
                     /label=5' LTR (truncated)
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7426..7650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7696..8124
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8138..8467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        8515..8562
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             8581..8952
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    9085..9218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(9303..9333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9348..9369)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9657..10245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10419..11276)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(11277..11381)
                     /label=AmpR promoter

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