LOCUS       sequence-463008-        5968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Ascl1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5968)
  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 5968)
  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: pMXs-mAscl1
FEATURES             Location/Qualifiers
     source          1..5968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    167..524
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             583..999
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     929..951
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     967..983
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1009..1383
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1321..1337
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    1426..1450
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1451..1460
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2153..2177)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2194..2782
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2247..2267)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2665..2676)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2962..3431
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3561..3578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3732..4320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3812..3831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4484..5275,5276..5344))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5107..5126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5345..5449)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(5479..5968,1..104)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
       61 tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg
      121 atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      181 cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt
      241 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttcggaa
      301 cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt tgtggcccga
      361 cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta ataggaggga
      421 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
      481 ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt
      541 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
      601 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
      661 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
      721 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
      781 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
      841 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
      901 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
      961 tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc
     1021 ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact
     1081 aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg
     1141 agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac
     1201 cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg
     1261 aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca
     1321 gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctaga
     1381 ctgccggatc tagctagtta atcgacggta tcgataagct atcaaacaag tttgtacaaa
     1441 aaagcaggct gccaccatgg agagctctgg caagatggag agtggagccg gccagcagcc
     1501 gcagcccccg cagcccttcc tgcctcccgc agcctgcttc tttgcgaccg cggcggcggc
     1561 ggcagcggcg gcggccgcgg cagctcagag cgcgcagcag caacagccgc aggcgccgcc
     1621 gcagcaggcg ccgcagctga gcccggtggc cgacagccag ccctcagggg gcggtcacaa
     1681 gtcagcggcc aagcaggtca agcgccagcg ctcgtcctct ccggaactga tgcgctgcaa
     1741 acgccggctc aacttcagcg gcttcggcta cagcctgcca cagcagcagc cggccgccgt
     1801 ggcgcgccgc aacgagcgcg agcgcaaccg ggtcaagttg gtcaacctgg gttttgccac
     1861 cctccgggag catgtcccca acggcgcggc caacaagaag atgagcaagg tggagacgct
     1921 gcgctcggcg gtcgagtaca tccgcgcgct gcagcagctg ctggacgagc acgacgcggt
     1981 gagcgctgcc tttcaggcgg gcgtcctgtc gcccaccatc tcccccaact actccaacga
     2041 cttgaactct atggcgggtt ctccggtctc gtcctactcc tccgacgagg gatcctacga
     2101 ccctcttagc ccagaggaac aagagctgct ggactttacc aactggttct gaacccagct
     2161 ttcttgtaca aagtggttcg atagcttatc gataatcaac ctctggatta caaaatttgt
     2221 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2281 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2341 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2401 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2461 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2521 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2581 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2641 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2701 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2761 tccctttggg ccgcctcccc gcatcgatac cgtcgacgat aaaataaaag attttattta
     2821 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     2881 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     2941 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     3001 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     3061 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     3121 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     3181 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     3241 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     3301 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     3361 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     3421 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     3481 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3541 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3601 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3661 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3721 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3781 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3841 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3901 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3961 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4021 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4081 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4141 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4201 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4261 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4321 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4381 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
     4441 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4621 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5341 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5401 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5461 gaaaagtgcc acctgcataa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     5521 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagaaaagt tcagatcaag
     5581 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     5641 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     5701 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     5761 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     5821 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     5881 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     5941 gattgactga gtcgcccggg tacccgtg
//
