pU6-Sp-pegRNA-HEK3_CTT_ins vector (V007064)

Basic Vector Information

      • Vector Name:
      • pU6-Sp-pegRNA-HEK3_CTT_ins
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 2305 bp
      • Type:
      • Mammalian Expression
      • Copy Number:
      • High Copy
      • Promoter:
      • U6
      • Cloning Method:
      • Ligation Independent Cloning
      • 5' Primer:
      • GACTATCATATGCTTACCGT

pU6-Sp-pegRNA-HEK3_CTT_ins vector Vector Map

pU6-Sp-pegRNA-HEK3_CTT_ins2305 bp60012001800gRNA scaffoldoriAmpRAmpR promoterU6 promoter

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 ℃.

pU6-Sp-pegRNA-HEK3_CTT_ins vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_44659        2305 bp DNA     circular SYN 13-MAY-2021
DEFINITION  S. pyogenes pegRNA for CTT insertion at the HEK3 site of human cells
            using prime editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2305)
  AUTHORS   Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, 
            Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR
  TITLE     Search-and-replace genome editing without double-strand breaks or 
            donor DNA.
  JOURNAL   Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 
            10.1038/s41586-019-1711-4.
  PUBMED    31634902
REFERENCE   2  (bases 1 to 2305)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2305)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 
            10.1038/s41586-019-1711-4."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        29..104
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(996..1853)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1854..1958)
                     /label=AmpR promoter
     promoter        2065..2305
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"

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