pX333-2×(U6-gRNA)-CBH-Cas9-Puro-EGFP vector (V017961)

Basic Vector Information

Vector Name:
pX333-2×(U6-gRNA)-CBH-Cas9-Puro-EGFP
Antibiotic Resistance:
Ampicillin
Length:
10439 bp
Type:
Gene-editing vector
Host:
Mammalian cells
Selection Marker:
Puro
Promoter:
U6
3' Primer:
BGH-R
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pX333-2×(U6-gRNA)-CBH-Cas9-Puro-EGFP vector Map

pX333-2×(U6-gRNA)-CBH-Cas9-Puro-EGFP10439 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000gRNA scaffoldU6 promoterCMV enhancerchicken β-actin promoterCas9EGFPbGH poly(A) signalAAV2 ITRf1 oriAmpR promoterAmpR

pX333-2×(U6-gRNA)-CBH-Cas9-Puro-EGFP vector Sequence

LOCUS       .                      10439 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_RNA        337..412
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        503..743
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        943..1228
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV
                     enhancer"
     promoter        1230..1507
                     /label="chicken β-actin promoter"
     CDS             1873..5973
                     /label="Cas9"
                     /note="S. pyogenes Cas9, codon optimized for C. elegans
                     with synthetic introns"
     CDS             6754..7470
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     polyA_signal    7504..7711
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7720..7860
                     /label="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
     rep_origin      7935..8390
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8672..8776
                     /label="AmpR promoter"
     CDS             8777..9634
                     /label="AmpR"
                     /note="β-lactamase"

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