LOCUS       sequence-307151-        3433 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fluorescent reporter for intestinal stem cells through ASCL2 
            transcriptional activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3433)
  AUTHORS   Heinz MC, Oost KC, Snippert HJG
  TITLE     Introducing the Stem Cell ASCL2 Reporter STAR into Intestinal 
            Organoids.
  JOURNAL   STAR Protoc. 2020 Oct 8;1(3):100126. doi: 
            10.1016/j.xpro.2020.100126. eCollection 2020 Dec 18.
  PUBMED    33377020
REFERENCE   2  (bases 1 to 3433)
  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: "STAR 
            Protoc."; date: "2020-10-8"; volume: "1(3):100126. doi"; pages: " 
            10.1016/j.xpro.2020.100126. eCollection 2020 Dec 18"
COMMENT     Sequence Label: Tol2 8xSTAR-sTomato. PGK-puro
FEATURES             Location/Qualifiers
     source          1..3433
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(962..980)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1230..1249
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1424..1447
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1569..1703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1781..2280
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1816..1839)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2213..2232
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2292..2339
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2388..2987
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2388..2407)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2884..2904
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3016..3150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3244..3406,3407..3418)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 agaggtgtaa agtacttgag taattttact tgattactgt acttaagtat tatttttggg
       61 gatttttact ttacttgagt acaattaaaa atcaatactt ttacttttac ttaattacat
      121 ttttttagaa aaaaaagtac tttttactcc ttacaatttt atttacagtc aaaaagtact
      181 tattttttgg agatcacttc attctatttt cccttgctat taccaaacca attgaattgc
      241 gctgatgccc agtttaattt aaatagatct atcgatcggt acccctaggg tgaaaaggca
      301 ggtgcaaagc aaatcctatc ccagcttcta gctaggtgaa aaggcaggtg caaagcaaat
      361 cctatcccag cttctagcta gggtgaaaag gcaggtgcaa agcaaatcct atcccagctt
      421 ctagctaggt gaaaaggcag gtgcaaagca aatcctatcc cagcttctag ctagggtgaa
      481 aaggcaggtg caaagcaaat cctatcccag cttctagcta ggtgaaaagg caggtgcaaa
      541 gcaaatccta tcccagcttc tagctagggt gaaaaggcag gtgcaaagca aatcctatcc
      601 cagcttctag ctaggtgaaa aggcaggtgc aaagcaaatc ctatcccagc ttctagctag
      661 cggcgcgccg gatccggagg ggacaagtgg agggaaagtg gggccgggcg gggggtgcct
      721 gggaagccag gctgcgctga cgtcactggg cgcgcaattc gggctggagc gctttaaaaa
      781 acgagcgtgc aagcagagag gctgctccac accgctcagg ccgcgaccat ggtgagcaag
      841 ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac
      901 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
      961 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     1021 cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag
     1081 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt
     1141 ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag
     1201 atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg
     1261 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag
     1321 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc
     1381 aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc
     1441 cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg
     1501 ttcctggggc atggcaccgg cagcaccggc agcggctaat ctagaactat agtgagtcgt
     1561 attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1621 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1681 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1741 agggggaggt gtgggaggtt ttttaattcg cggccgcacc gggtagggga ggcgcttttc
     1801 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     1861 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     1921 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     1981 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     2041 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     2101 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     2161 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     2221 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     2281 acctgcagcc tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     2341 aggtgaggaa ctaaaccgtc gacgatttca ctagtctcga agccaccatg accgagtaca
     2401 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     2461 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     2521 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     2581 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     2641 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     2701 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     2761 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     2821 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     2881 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     2941 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacta gaactatagt
     3001 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     3061 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3121 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3181 caggttcagg gggaggtgtg ggaggttttt taattaagct taaacaagaa tctctagttt
     3241 tctttcttgc ttttactttt acttccttaa tactcaagta caattttaat ggagtacttt
     3301 tttactttta ctcaagtaag attctagcca gatactttta cttttaattg agtaaaattt
     3361 tccctaagta cttgtacttt cacttgagta aaatttttga gtacttttta cacctctgct
     3421 cgaccatatg gga
//
