ptCC9 vector (V002803)

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