LOCUS       sequence-463825-        4117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Isl1 with mRuby2 fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4117)
  AUTHORS   Wang NB, Lende-Dorn BA, Beitz AM, Han P, Adewumi HO, O'Shea TM, 
            Galloway KE
  TITLE     Proliferation history and transcription factor levels drive direct 
            conversion to motor neurons.
  JOURNAL   Cell Syst. 2025 Apr 16;16(4):101205. doi: 
            10.1016/j.cels.2025.101205. Epub 2025 Mar 13.
  PUBMED    40086434
REFERENCE   2  (bases 1 to 4117)
  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: "Cell 
            Syst."; date: "2025-04-16"; volume: "16(4):101205. doi"; pages: " 
            10.1016/j.cels.2025.101205. Epub 2025 Mar 13"
COMMENT     Sequence Label: pENTR-Isl1-P2A-mRuby2
FEATURES             Location/Qualifiers
     source          1..4117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(14..113)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(225..244)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             236..1045
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(303..322)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1135..1723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1624..1643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1888..1974
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2066..2093
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2101..2124
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    2143..2242
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2285..2294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3081..3090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3347..3403
                     /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             3404..4114
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
ORIGIN
        1 cgctgatatc tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
       61 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
      121 gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
      181 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
      241 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
      301 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
      361 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
      421 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
      481 caagcatttt atccgtactc ctggtgatgc atggttactc accactgcga tccccggaaa
      541 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
      601 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
      661 tcgcgtattt cgtctagctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
      721 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
      781 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
      841 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
      901 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
      961 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     1021 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac attattcaga
     1081 ttgggccccg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1141 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1201 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1261 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1321 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1381 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1441 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1501 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1561 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1621 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1681 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1741 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1801 ccctgattct gtggataacc gtattaccgc tagcatggat ctcggggacg tctaactact
     1861 aagcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg
     1921 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1981 ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     2041 taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt
     2101 ctacaaactc ttcctgttag ttagttactt aagctcgggc cccaaataat gattttattt
     2161 tgactgatag tgacctgttc gttgcaacaa attgataagc aatgcttttt tataatgcca
     2221 actttgtaca aaaaagcagg ctccaccatg ggaaccaatt cagtcgactg gatccggtac
     2281 tactgccacc atgggagaca tgggcgatcc accaaaaaaa aaacgtctga tttccctgtg
     2341 tgttggttgc ggcaatcaaa ttcacgacca gtatattctg agggtttctc cggatttgga
     2401 gtggcatgca gcatgtttga aatgtgcgga gtgtaatcag tatttggacg aaagctgtac
     2461 gtgctttgtt agggatggga aaacctactg taaaagagat tatatcaggt tgtacgggat
     2521 caaatgcgcc aagtgcagca taggcttcag caagaacgac ttcgtgatgc gcgcccgctc
     2581 taaggtgtac cacatcgagt gtttccgctg tgtagcctgc agccgacagc tcatcccggg
     2641 agacgaattc gccctgcggg aggatgggct tttctgccgt gcagaccacg atgtggtgga
     2701 gagagccagc ctgggagctg gagatcctct cagtcccttg catccagcgc ggcctctgca
     2761 aatggcagcc gaacccatct cggctaggca gccagctctg cggccgcacg tccacaagca
     2821 gccggagaag accacccgag tgcggactgt gctcaacgag aagcagctgc acaccttgcg
     2881 gacctgctat gccgccaacc ctcggccaga tgcgctcatg aaggagcaac tagtggagat
     2941 gacgggcctc agtcccagag tcatccgagt gtggtttcaa aacaagcggt gcaaggacaa
     3001 gaaacgcagc atcatgatga agcagctcca gcagcagcaa cccaacgaca aaactaatat
     3061 ccaggggatg acaggaactc ccatggtggc tgctagtccg gagagacatg atggtggttt
     3121 acaggctaac ccagtagagg tgcaaagtta ccagccgccc tggaaagtac tgagtgactt
     3181 cgccttgcaa agcgacatag atcagcctgc ttttcagcaa ctggtcaatt tttcagaagg
     3241 aggaccaggc tctaattcta ctggcagtga agtagcatcg atgtcctcgc agctcccaga
     3301 tacacccaac agcatggtag ccagtcctat tgaggcagga agcggagcta ctaacttcag
     3361 cctgctgaag caggctggag acgtggagga gaaccctgga cctatggtgt ctaagggcga
     3421 agagctgatc aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca
     3481 ccaattcaaa tgcacaggtg aaggagaagg caatccgtac atgggaactc aaaccatgag
     3541 gatcaaagtc atcgagggag gacccctgcc atttgccttt gacattcttg ccacgtcgtt
     3601 catgtatggc agccgtactt ttatcaagta cccgaaaggc attcctgatt tctttaaaca
     3661 gtcctttcct gagggtttta cttgggaaag agttacgaga tacgaagatg gtggagtcgt
     3721 caccgtcatg caggacacca gccttgagga tggctgtctc gtttaccacg tccaagtcag
     3781 aggggtaaac tttccctcca atggtcccgt gatgcagaag aagaccaagg gttgggagcc
     3841 taatacagag atgatgtatc cagcagatgg tggtctgagg ggatacactc atatggcact
     3901 gaaagttgat ggtggtggcc atctgtcttg ctctttcgta acaacttaca ggtcaaaaaa
     3961 gaccgtcggg aacatcaaga tgcccggtat ccatgccgtt gatcaccgcc tggaaaggtt
     4021 agaggaaagt gacaatgaaa tgttcgtagt acaacgcgaa cacgcagttg ccaagttcgc
     4081 cgggcttggt ggtgggatgg acgagctgta caagtaa
//
