LOCUS       sequence-122243-         818 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A Sleeping beauty transposon with conditional expressed hCas9. A red
            flourescent gene linked to puromycin can be removed in the prescence
            of Flp recombinase allowing Cas9 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 818)
  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 818)
  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: LIR_TRE_CAG_mKateII-Puro-STOP_mVenus-Cas9-PA-RIR
FEATURES             Location/Qualifiers
     source          1..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    2..49
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      50..59
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             56..751
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
ORIGIN
        1 agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcg ccaccatggt
       61 gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa
      121 ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
      181 gagaatcaag gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag
      241 cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa
      301 gcagtccttc cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
      361 gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat
      421 cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga
      481 ggcctccacc gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc
      541 cctgaagctc gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa
      601 gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag
      661 aatcaaggag gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata
      721 ctgcgacctc cctagcaaac tggggcacag acctagggga agcggacagt gtactaatta
      781 tgctctcttg aaattggctg gagatgttga gagcaacc
//
