LOCUS       sequence-307147-        2114 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 2114)
  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 2114)
  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: pLV 4xSTAR-mScarletI-NLS-blast
FEATURES             Location/Qualifiers
     source          1..2114
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             365..1060
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(498..516)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     766..785
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1067..1087
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1137..1689
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1281..1298)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1508..1527
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1710..1719
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1716..2114
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 gtgaaaaggc aggtgcaaag caaatcctat cccagcttct agctaggtga aaaggcaggt
       61 gcaaagcaaa tcctatccca gcttctagct agggtgaaaa ggcaggtgca aagcaaatcc
      121 tatcccagct tctagctagg tgaaaaggca ggtgcaaagc aaatcctatc ccagcttcta
      181 gctagcggcg cgccggatcc ggaggggaca agtggaggga aagtggggcc gggcgggggg
      241 tgcctgggaa gccaggctgc gctgacgtca ctgggcgcgc aattcgggct ggagcgcttt
      301 aaaaaacgag cgtgcaagca gagaggctgc tccacaccgc tcaggccgcg gtttaaacat
      361 cgatatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca aggtgcacat
      421 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
      481 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttctcctg
      541 ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcatcaagc accccgccga
      601 catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa
      661 cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg acggcaccct
      721 gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg taatgcagaa
      781 gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa
      841 gggcgacatt aagatggccc tgcgcctgaa ggacggcgga cgctacctgg cggacttcaa
      901 gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg tggaccgcaa
      961 gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg aacgctccga
     1021 gggccgccac tccaccggcg gcatggacga gctgtacaag tccaggccaa aaaagaaaag
     1081 aaaggtcatc gattagttaa ttaatggccc gggaattccg cccctctccc cccctaacgt
     1141 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     1201 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     1261 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     1321 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     1381 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     1441 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     1501 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     1561 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     1621 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     1681 atgataatat ggatatggcc acaacggtcg ccaccatggc caagcctttg tctcaagaag
     1741 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     1801 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     1861 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     1921 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     1981 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     2041 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     2101 tgtgggaggg ctaa
//
