LOCUS       sequence-98925-e        3515 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid mediates polycistronic expression of 4 genes: Ngn3, 
            Pdx1, Mafa, and mCherry. It can be used to convert mouse pancreatic 
            acinar cells to induced beta-like cells in vivo. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3515)
  AUTHORS   Li W, Cavelti-Weder C, Zhang Y, Clement K, Donovan S, Gonzalez G, 
            Zhu J, Stemann M, Xu K, Hashimoto T, Yamada T, Nakanishi M, Zhang Y,
            Zeng S, Gifford D, Meissner A, Weir G, Zhou Q
  TITLE     Long-term persistence and development of induced pancreatic beta 
            cells generated by lineage conversion of acinar cells.
  JOURNAL   Nat Biotechnol. 2014 Dec;32(12):1223-30. doi: 10.1038/nbt.3082. Epub
            2014 Nov 17.
  PUBMED    25402613
REFERENCE   2  (bases 1 to 3515)
  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"; doi: "10.1038/nbt.3082"; 
            journalName: "Nat Biotechnol"; date: "2014-12"; volume: "32"; issue:
            "12"; pages: "1223-30"
COMMENT     Sequence Label: pAd-M3cherry
FEATURES             Location/Qualifiers
     source          1..3515
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      10..19
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             664..720
                     /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             1585..1638
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2233..2262
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2728..2787
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             2797..3507
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2942..2960)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3210..3229
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gtcgacgccg ccaccatggc gcctcatccc ttggatgcgc tcaccatcca agtgtcccca
       61 gagacacaac aaccttttcc cggagcctcg gaccacgaag tgctcagttc caattccacc
      121 ccacctagcc ccactctcat acctagggac tgctccgaag cagaagtggg tgactgccga
      181 gggacctcga ggaagctccg cgcccgacgc ggagggcgca acaggcccaa gagcgagttg
      241 gcactcagca aacagcgaag aagccggcgc aagaaggcca atgatcggga gcgcaatcgc
      301 atgcacaacc tcaactcggc gctggatgcg ctgcgcggtg tcctgcccac cttcccggat
      361 gacgccaaac ttacaaagat cgagaccctg cgcttcgccc acaactacat ctgggcactg
      421 actcagacgc tgcgcatagc ggaccacagc ttctatggcc cggagccccc tgtgccctgt
      481 ggagagctgg ggagccccgg aggtggctcc aacggggact ggggctctat ctactcccca
      541 gtctcccaag cgggtaacct gagccccacg gcctcattgg aggaattccc tggcctgcag
      601 gtgcccagct ccccatccta tctgctcccg ggagcactgg tgttctcaga cttcttgact
      661 agtgccacga acttctctct gttaaagcaa gcaggagatg ttgaagaaaa ccccgggcct
      721 ggatccatga acagtgagga gcagtactac gcggccacac agctctacaa ggacccgtgc
      781 gcattccaga ggggcccggt gccagagttc agcgctaacc cccctgcgtg cctgtacatg
      841 ggccgccagc ccccacctcc gccgccaccc cagtttacaa gctcgctggg atcactggag
      901 cagggaagtc ctccggacat ctccccatac gaagtgcccc cgctcgcctc cgacgacccg
      961 gctggcgctc acctccacca ccaccttcca gctcagctcg ggctcgccca tccacctccc
     1021 ggacctttcc cgaatggaac cgagcctggg ggcctggaag agcccaaccg cgtccagctc
     1081 cctttcccgt ggatgaaatc caccaaagct cacgcgtgga aaggccagtg ggcaggaggt
     1141 gcttacacag cggaacccga ggaaaacaag aggacccgta ctgcctacac ccgggcgcag
     1201 ctgctggagc tggagaagga attcttattt aacaaataca tctcccggcc ccgccgggtg
     1261 gagctggcag tgatgttgaa cttgaccgag agacacatca aaatctggtt ccaaaaccgt
     1321 cgcatgaagt ggaaaaaaga ggaagataag aaacgtagta gcgggacccc gagtgggggc
     1381 ggtgggggcg aagagccgga gcaagattgt gcggtgacct cgggcgagga gctgctggca
     1441 gtgccaccgc tgccacctcc cggaggtgcc gtgcccccag gcgtcccagc tgcagtccgg
     1501 gagggcctac tgccttcggg ccttagcgtg tcgccacagc cctccagcat cgcgccactg
     1561 cgaccgcagg aaccccgggg atccgagggc agaggaagtc ttctaacatg cggtgacgtg
     1621 gaggagaatc ccggccctat cgatatggcc gcggagctgg cgatgggcgc agagctgccc
     1681 agcagcccac tggccatcga gtacgtcaac gacttcgacc tgatgaagtt cgaggtgaag
     1741 aaggagccgc ccgaggccga gcgcttctgc caccgcctgc cgcccggctc gctgtcctcg
     1801 acgcccctca gcacgccctg ctcctcggtg ccctcttcgc ccagcttctg cgcacccagc
     1861 ccgggcacag gcggcggcgc gggcggcggg ggcagcgcgg ctcaggccgg gggcgccccg
     1921 gggccgccga gtggaggccc cggcactgtc gggggcgcct caggaaaagc ggtgctggag
     1981 gatctgtact ggatgagcgg gtaccagcac cacctgaacc ccgaggcgct caacctgacg
     2041 ccggaggacg cggtggaggc gctcatcggc agcggccacc acggcgcgca ccacggcgcg
     2101 catcacccgg cggctgctgc ggcctatgag gccttccggg gtcagagctt cgcgggcggc
     2161 ggcggcgcgg acgacatggg tgccggccac caccacggcg cacaccacac tgcccaccat
     2221 catcactctg cccaccatca ccatcaccac catcaccacc acggaggctc tggccaccac
     2281 ggcggaggcg cgggtcacgg cggaggcggc gcaggccacc acgtgcgctt ggaggagcgc
     2341 ttctccgacg accagctggt atccatgtcc gtgcgggagc tgaaccggca gctccgcggc
     2401 ttcagcaagg aggaggtcat ccgactgaaa cagaagcggc gcacgctcaa gaaccgcggc
     2461 tacgcgcagt cgtgccgctt caagcgggtg cagcagcggc acattctgga gagcgagaag
     2521 tgccagctcc agagccaggt ggagcagctg aagctggagg tggggcgtct ggccaaggag
     2581 cgggacctgt acaaggagaa atacgagaag ttggcgggcc ggggcggccc cgggggcgcg
     2641 ggcggggccg gcttccctcg ggagccctcg ccagcgcagg ctggccccgg ggcggccaaa
     2701 ggcgcacccg acttctttct gatcgatcag tgtactaatt atgctctctt gaaattggct
     2761 ggagatgttg agagcaaccc aggtcccgcg gccgcaatgg tgagcaaggg cgaggaggat
     2821 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     2881 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2941 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3001 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3061 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3121 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3181 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3241 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3301 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3361 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3421 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3481 ggcggcatgg acgagctgta caagtaagcg gccgc
//
