P1-EYFP-Triplex-28-gRNA2-28-pA vector (V009797)

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 Map

P1-EYFP-Triplex-28-gRNA2-28-pA4592 bp600120018002400300036004200oriAmpRAmpR promoterf1 oripause site8x gRNA1 binding sitesMinimal Ade MLPPhiYFPMALAT1_3 triplexCsy4 rptgRNA scaffoldCsy4 rptSV40 poly(A) signal

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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