LOCUS       sequence-122835-         788 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 788)
  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 788)
  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..788
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(310..330)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(337..402)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(403..459)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(469..720)
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(511..532)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt
       61 cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc
      121 gccgctgtcg aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt
      181 gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc
      241 cacagagttg gtgccgatgt ccaggccgat gctgtacttc ttgtcggctg ctgggactcc
      301 gtggataccg accttccgct tcttctttgg ggccatctta tcgtcatcgt ctttgtaatc
      361 aatatcatga tccttgtagt ctccgtcgtg gtccttatag tcaggtccag ggttctcctc
      421 cacgtctcca gcctgcttca gcaggctgaa gttagtagct ccgcttccct tgtacagctc
      481 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
      541 cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt tgtcgggcag
      601 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc
      661 gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc
      721 ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
      781 gccgtcct
//
