Basic Vector Information
MS2-P65-HSF1_GFP 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 ℃.
MS2-P65-HSF1_GFP vector Sequence
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
This page is informational only.