LOCUS       sequence-149702-         854 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 codon optimized for Phaeodactylum tricornutum and a 
            sgRNA driven by a U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 854)
  AUTHORS   Nymark M, Sharma AK, Sparstad T, Bones AM, Winge P
  TITLE     A CRISPR/Cas9 system adapted for gene editing in marine algae.
  JOURNAL   Sci Rep. 2016 Apr 25;6:24951. doi: 10.1038/srep24951.
  PUBMED    27108533
REFERENCE   2  (bases 1 to 854)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-04-25"; volume: "6:24951. doi"; pages: " 
            10.1038/srep24951"
COMMENT     Sequence Label: pKS diaCas9_sgRNA
FEATURES             Location/Qualifiers
     source          1..854
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        223..241
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     269..285
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     270..286
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(633..708)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag
      241 ggaacaaaag ctggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgcagaa
      301 aagttcgtcg agcccatggc gattttagat tcgagatctt tgaaaattct gctaatcacc
      361 agcctcctga aataagaatg ttatagatag ccaattccgc tcactcacac tcgtgttcca
      421 aactgtaaac gcacttgtcc ataaattctc tcgaactcgc aaaagccaag acatgctacc
      481 cgcagaccgg tcggtagtaa agcaaccgga aaacaattta gtcaaacgaa atgtctaggg
      541 cttcagcgag ctgaagttat cgcaaacgca ttcatagcag tttgtcaatg taaatgacat
      601 acgatagcat gggtgagcgg ttctaaaaaa aagcaccgac tcggtgccac tttttcaagt
      661 tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaactg agaccctctc
      721 tcggtctcct cgactttgaa ggtgtttttt gaccttataa agctctaaaa tattgttgac
      781 taacaatgaa tacttggcgt gtcaaagtac cttgtagata ttatagatcc tattcaattt
      841 accgtcaatt cgac
//
