LOCUS       sequence-41531-e         880 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 880)
  AUTHORS   Koo BK, Stange DE, Sato T, Karthaus W, Farin HF, Huch M, van Es JH, 
            Clevers H
  TITLE     Controlled gene expression in primary Lgr5 organoid cultures.
  JOURNAL   Nat Methods. 2011 Dec 4. doi: 10.1038/nmeth.1802.
  PUBMED    22138822
REFERENCE   2  (bases 1 to 880)
  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: "Nat 
            Methods. 2011 Dec 4. doi: 10.1038/nmeth.1802."
COMMENT     Sequence Label: pMSCV-loxp-dsRed-loxp-3xHA-Puro-WPRE
FEATURES             Location/Qualifiers
     source          1..880
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(53..86)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      115..124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             121..798
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(301..321)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     519..538
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     713..736
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(823..856)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 actccttctc taggcgccgg aaattagatc actagtccag tgtggtggaa ttataacttc
       61 gtatagcata cattatacga agttataatt ctgcagatat ccagcacagt ggcggccacc
      121 atggatagca ctgagaacgt catcaagccc ttcatgcgct tcaaggtgca catggagggc
      181 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggcaagcc ctacgagggc
      241 acccagaccg ccaagctgca ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
      301 ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc cgacatcccc
      361 gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      421 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcac cttcatctac
      481 cacgtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca gaagaagact
      541 ctgggctggg agccctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
      601 atccacaagg cgctgaagct gaagggcggc ggccactacc tggtggagtt caagtcaatc
      661 tacatggcca agaagcccgt gaagctgccc ggctactact acgtggactc caagctggac
      721 atcacctccc acaacgagga ctacaccgtg gtggagcagt acgagcgcgc cgaggcccgc
      781 caccacctgt tccagtagag ctagctgaat aaggccgctc gaataacttc gtatagcata
      841 cattatacga agttattcga gtctagaggg cccgttagat
//
