ptCC9 vector (V002803)

Basic Vector Information

      • Vector Name:
      • ptCC9
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 8857 bp
      • Type:
      • Shuttle vector
      • Source/Author:
      • Stukenberg D, Zauner S, Dell'Aquila G, Maier UG.

ptCC9 vector Vector Map

ptCC98857 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800nitrate reductase; derived from Phaeodactylum tricornutum3xFLAGSV40 NLSCas9nucleoplasmin NLSnitrate reductase; derived from Phaeodactylum tricornutumU6-RNA promoter; derived from Phaeodactylum tricornutumgRNA scaffoldU6-RNA terminator; derived from Phaeodactylum tricornutumfcpB promoter; derived from Phaeodactylum tricornutumBleoRfcpB teminator; derived from Phaeodactylum tricornutumT7 promoteroriAmpRAmpR 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 ℃.

ptCC9 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_42714        8857 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector ptCC9, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8857)
  AUTHORS   Stukenberg D, Zauner S, Dell'Aquila G, Maier UG.
  TITLE     Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum
  JOURNAL   Front Plant Sci 9, 740 (2018)
  PUBMED    29928285
REFERENCE   2  (bases 1 to 8857)
  AUTHORS   Stuckenberg D, Zauner S.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2018) Philipps-University Marburg, Cellbiology, 
            Karl-von-Frisch Str. 8, Marburg 35043, Deutschland
REFERENCE   3  (bases 1 to 8857)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8857)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Plant
            Sci 9, 740 (2018)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2018) Philipps-University Marburg, Cellbiology, 
            Karl-von-Frisch Str. 8, Marburg 35043, Deutschland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..415
                     /note="nitrate reductase; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="promoter"
     CDS             431..496
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             503..523
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             548..4648
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             4649..4696
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     regulatory      4700..4972
                     /note="nitrate reductase; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="terminator"
     regulatory      5010..5288
                     /note="U6-RNA promoter; derived from Phaeodactylum
                     tricornutum"
                     /regulatory_class="promoter"
     misc_RNA        5310..5385
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      5386..5723
                     /note="U6-RNA terminator; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="terminator"
     regulatory      5724..5916
                     /note="fcpB promoter; derived from Phaeodactylum
                     tricornutum"
                     /regulatory_class="promoter"
     CDS             5917..6288
                     /label=BleoR
                     /note="antibiotic-binding protein"
     regulatory      6292..6532
                     /note="fcpB teminator; derived from Phaeodactylum
                     tricornutum"
                     /regulatory_class="terminator"
     promoter        complement(6543..6561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(6819..7407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7581..8438)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8439..8543)
                     /label=AmpR promoter

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