LOCUS       sequence-131575-         610 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Used to produce AAV expressing the FlpO recombinase and pig gRNA 
            towards 3 tumor suppressors: PTEN, p53 and SMAD4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 610)
  AUTHORS   Berthelsen MF, Riedel M, Cai H, Skaarup SH, Alstrup AKO, 
            Dagnaes-Hansen F, Luo Y, Jensen UB, Hager H, Liu Y, Callesen H, 
            Vendelbo MH, Jakobsen JE, Thomsen MK
  TITLE     The CRISPR/Cas9 Minipig-A Transgenic Minipig to Produce Specific 
            Mutations in Designated Tissues.
  JOURNAL   Cancers (Basel). 2021 Jun 16;13(12). pii: cancers13123024. doi: 
            10.3390/cancers13123024.
  PUBMED    34208747
REFERENCE   2  (bases 1 to 610)
  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: "Cancers 
            (Basel)."; date: "2021-06-16"; volume: "13(12). pii: 
            cancers13123024. doi"; pages: " 10.3390/cancers13123024"
COMMENT     Sequence Label: AAV_3xgRNA;PTENA, p53B,SMAD4A_CAG_FlPO_synthPA
FEATURES             Location/Qualifiers
     source          1..610
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(114..189)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(210..458)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(268..287)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(438..458)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_RNA        complement(471..546)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gacgtcaatg gaaagtccct attggcgtta ctattgacgt caatgggcgg gggtcgttgg
       61 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gggtaccaaa aaagcaccga
      121 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
      181 ctctaaaaca ttcatctttt ttctccttcg gtgtttcgtc ctttccacaa gatatataaa
      241 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
      301 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
      361 aatatctttg tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt
      421 atatgcaaat atgaaggaat catgggaaat aggccctcat cgataaaaaa gcaccgactc
      481 ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta tttctagctc
      541 taaaacggac aggcacaaac acgcacggtg tttcgtcctt tccacaagat atataaagcc
      601 aagaaatcga
//
