gRNA_GFP-T1 vector (V012340)

LOCUS       gRNA_GFP-T1.        3973 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid for guide RNA (gRNA) targeting of an integrated GFP gene in 
            human cells using the GFP T1 target sequence.
ACCESSION   .
VERSION     .
KEYWORDS    gRNA_GFP-T1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3973)
  AUTHORS   Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE,
            Church GM.
  TITLE     RNA-guided human genome engineering via Cas9.
  JOURNAL   Science 2013;339:823-6.
  PUBMED    23287722
REFERENCE   2  (bases 1 to 3973)
  AUTHORS   Church Lab / Addgene #41819
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3973)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "2013"; volume: "339"; pages: "823-6"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     terminator      372..377
                     /note="polIII terminator"
                     /note="RNA polymerase III transcription terminator"
     misc_RNA        complement(378..453)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    454..473
                     /label=GFP T1
                     /note="GFP T1"
                     /note="GFP gRNA Target 1 sequence"
     promoter        complement(482..722)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        complement(862..880)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(887..903)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1041..1340
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             1692..2483
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             2692..3063
                     /label=BleoR
                     /note="antibiotic-binding protein"
     rep_origin      3204..3792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"