pTargetF vector (V012144)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012144 pTargetF In stock (lyophilized plasmid)

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Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.

Basic Vector Information

The pTargetF is for constitutive expression of sgRNA without donor editing template DNA.

      • Vector Name:
      • pTargetF
      • Antibiotic Resistance:
      • Spectinomycin, 50 μg/mL
      • Length:
      • 2117 bp
      • Type:
      • Bacterial Expression, CRISPR
      • Copy Number:
      • High Copy
      • Promoter:
      • pij23119
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • MoClo-F (agcgaggaagcggaagagcg)
      • Growth Strain(s):
      • DH5alpha
      • Growth Temperature:
      • 37℃

pTargetF vector Vector Map

pTargetF2117 bp60012001800L4440J23119(SpeI) promotergRNA scaffoldSmRori

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 ℃.

References

  • Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang S. Multigene editing in the Escherichia coli genome via the CRISPR-Cas9 system. Appl Environ Microbiol. 2015 Apr;81(7):2506-14. doi: 10.1128/AEM.04023-14. Epub 2015 Jan 30. Erratum in: Appl Environ Microbiol. 2016 Jun 15;82(12):3693.

pTargetF vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_42689        2117 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Constitutive expression of sgRNA without donor editing template DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2117)
  AUTHORS   Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang S
  TITLE     Multigene editing in the Escherichia coli genome using the 
            CRISPR-Cas9 system.
  JOURNAL   Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14.
  PUBMED    25636838
REFERENCE   2  (bases 1 to 2117)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2117)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        192..226
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        247..322
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             508..1296
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1473..2061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"