Basic Vector Information
- Vector Name:
- P1-EYFP-Triplex-28-gRNA2-28-pA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4592 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.
P1-EYFP-Triplex-28-gRNA2-28-pA 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 ℃.
P1-EYFP-Triplex-28-gRNA2-28-pA vector Sequence
LOCUS 40924_2353 4592 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector P1-EYFP-Triplex-28-gRNA2-28-pA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4592)
AUTHORS Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.
TITLE Multiplexed and programmable regulation of gene networks with an
integrated RNA and CRISPR/Cas toolkit in human cells
JOURNAL Mol. Cell 54 (4), 698-710 (2014)
PUBMED 24837679
REFERENCE 2 (bases 1 to 4592)
AUTHORS Perli SD.
TITLE Direct Submission
JOURNAL Submitted (04-MAY-2014) Electrical Engineering and Computer Science,
Massachusetts Institute of Technology, 77, Massachusetts Avenue,
Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 4592)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4592)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Cell";
date: "2014"; volume: "54"; issue: "4"; pages: "698-710"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-MAY-2014) Electrical Engineering and Computer Science,
Massachusetts Institute of Technology, 77, Massachusetts Avenue,
Cambridge, MA 02139, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4592
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(99..687)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(861..1718)
/label=AmpR
/note="beta-lactamase"
promoter complement(1719..1823)
/label=AmpR promoter
rep_origin 1850..2305
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
misc_feature 2496..2587
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
misc_feature 2601..2962
/label=8x gRNA1 binding sites
/note="8x gRNA1 binding sites"
regulatory 2969..3030
/label=Minimal Ade MLP
/note="Minimal Ade MLP"
/regulatory_class="promoter"
CDS 3037..3738
/label=PhiYFP
/note="Phialidium yellow fluorescent proteim"
misc_feature 3751..3860
/label=MALAT1_3 triplex
/note="MALAT1_3 triplex"
misc_feature 3911..3938
/label=Csy4 rpt
/note="Csy4 rpt"
protein_bind 3916..3930
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 3959..4034
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 4043..4070
/label=Csy4 rpt
/note="Csy4 rpt"
protein_bind 4048..4062
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
polyA_signal complement(4146..4267)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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