LOCUS       sequence-307149-        2126 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 2126)
  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 2126)
  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-mNeonGreen-NLS-blast
FEATURES             Location/Qualifiers
     source          1..2126
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             365..1072
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      620..629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1079..1099
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1149..1701
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1293..1310)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1520..1539
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1722..1731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1728..2126
                     /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 aggaggataa catggcctct ctcccagcga cacatgagtt
      421 acacatcttt ggctccatca acggtgtgga ctttgacatg gtgggtcagg gcaccggcaa
      481 tccaaatgat ggttatgagg agttaaacct gaagtccacc aagggtgacc tccagttctc
      541 cccctggatt ctggtccctc atatcgggta tggcttccat cagtacctgc cctaccctga
      601 cgggatgtcg cctttccagg ccgccatggt agatggctcc ggataccaag tccatcgcac
      661 aatgcagttt gaagatggtg cctcccttac tgttaactac cgctacacct acgagggaag
      721 ccacatcaaa ggagaggccc aggtgaaggg gactggtttc cctgctgacg gtcctgtgat
      781 gaccaactcg ctgaccgctg cggactggtg caggtcgaag aagacttacc ccaacgacaa
      841 aaccatcatc agtaccttta agtggagtta caccactgga aatggcaagc gctaccggag
      901 cactgcgcgg accacctaca cctttgccaa gccaatggcg gctaactatc tgaagaacca
      961 gccgatgtac gtgttccgta agacggagct caagcactcc aagaccgagc tcaacttcaa
     1021 ggagtggcaa aaggccttta ccgatgtgat gggcatggac gagctgtaca agtccaggcc
     1081 aaaaaagaaa agaaaggtca tcgattagtt aattaatggc ccgggaattc cgcccctctc
     1141 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1201 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1261 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1321 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     1381 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     1441 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     1501 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     1561 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     1621 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     1681 ttgaaaaaca cgatgataat atggatatgg ccacaacggt cgccaccatg gccaagcctt
     1741 tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca
     1801 tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg
     1861 gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg
     1921 ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg
     1981 gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca
     2041 aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc
     2101 cctctggtta tgtgtgggag ggctaa
//
