LOCUS       sequence-465028-        8495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Ngn2, Isl1, and Lhx3 for motor neuron 
            reprogramming.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8495)
  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 8495)
  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-NIL
FEATURES             Location/Qualifiers
     source          1..8495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..130
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    219..243
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1090..1146
                     /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"
     regulatory      1937..1946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2203..2256
                     /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"
     protein_bind    complement(3473..3497)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3514..4102
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3567..3587)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3985..3996)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4282..4751
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4881..4898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5052..5640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5132..5151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5804..6595,6596..6664))
                     /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     6427..6446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6665..6769)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6799..7392
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7455..7812
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7871..8287
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8217..8239
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8255..8271
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(8297..8495,1..176)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 gcgacacagt gtgggtccgc cgacaccaga ctaagaacct agaacctcgc tggaaaggac
       61 cttacacagt cctgctgacc acccccaccg ccctcaaagt agacggcatc gcagcttgga
      121 tacacgccgc ccacgtgaag gctgccgacc ccgggggtgg accatcctct agactgccgg
      181 atctagctag ttaatcgacg gtatcgataa gctatcaaac aagtttgtac aaaaaagcag
      241 gctccaccat gggaaccaat tcagtcgact ggatccggta ctactgccac catgttcgtc
      301 aaatctgaga ctctggagtt gaaggaggaa gaggaggtac tgatgctgct gggctcggct
      361 tccccggcct cggcgaccct gaccccgatg tcctccagcg cggacgagga ggaggacgag
      421 gagctgcgcc ggccgggctc cgcgcgtggg cagcgtggag cggaagccgg gcagggggtg
      481 cagggcagtc cggcgtcggg tgccgggggt tgccggccag ggcggctgct gggcctgatg
      541 cacgagtgca agcgtcgccc gtcgcgctca cgggccgtct cccgaggtgc caagacggcg
      601 gagacggtgc agcgcatcaa gaagacccgc aggctcaagg ccaacaaccg cgagcgcaac
      661 cgcatgcaca acctaaacgc cgcgctggac gcgctgcgcg aggtgctgcc caccttcccc
      721 gaggatgcca agctcacgaa gatcgagacg ctgcgcttcg cccacaatta catctgggcg
      781 ctcaccgaga ctctgcgcct ggcggaccac tgcgccggcg ccggtggcct ccagggggcg
      841 ctcttcacgg aggcggtgct cctgagcccg ggagctgcgc tcggcgccag cggggacagc
      901 ccttctccac cttcctcctg gagctgcacc aacagcccgg cgtcatcctc caactccacg
      961 tccccataca gctgcacttt atcgcccgct agccccgggt cagacgtgga ctactggcag
     1021 cccccacctc cggagaagca tcgttatgcg cctcacctgc ccctcgccag ggactgtatc
     1081 ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct
     1141 ggacctatgg gagacatggg cgatccacca aaaaaaaaac gtctgatttc cctgtgtgtt
     1201 ggttgcggca atcaaattca cgaccagtat attctgaggg tttctccgga tttggagtgg
     1261 catgcagcat gtttgaaatg tgcggagtgt aatcagtatt tggacgaaag ctgtacgtgc
     1321 tttgttaggg atgggaaaac ctactgtaaa agagattata tcaggttgta cgggatcaaa
     1381 tgcgccaagt gcagcatagg cttcagcaag aacgacttcg tgatgcgcgc ccgctctaag
     1441 gtgtaccaca tcgagtgttt ccgctgtgta gcctgcagcc gacagctcat cccgggagac
     1501 gaattcgccc tgcgggagga tgggcttttc tgccgtgcag accacgatgt ggtggagaga
     1561 gccagcctgg gagctggaga tcctctcagt cccttgcatc cagcgcggcc tctgcaaatg
     1621 gcagccgaac ccatctcggc taggcagcca gctctgcggc cgcacgtcca caagcagccg
     1681 gagaagacca cccgagtgcg gactgtgctc aacgagaagc agctgcacac cttgcggacc
     1741 tgctatgccg ccaaccctcg gccagatgcg ctcatgaagg agcaactagt ggagatgacg
     1801 ggcctcagtc ccagagtcat ccgagtgtgg tttcaaaaca agcggtgcaa ggacaagaaa
     1861 cgcagcatca tgatgaagca gctccagcag cagcaaccca acgacaaaac taatatccag
     1921 gggatgacag gaactcccat ggtggctgct agtccggaga gacatgatgg tggtttacag
     1981 gctaacccag tagaggtgca aagttaccag ccgccctgga aagtactgag tgacttcgcc
     2041 ttgcaaagcg acatagatca gcctgctttt cagcaactgg tcaatttttc agaaggagga
     2101 ccaggctcta attctactgg cagtgaagta gcatcgatgt cctcgcagct cccagataca
     2161 cccaacagca tggtagccag tcctattgag gcaggcagtg gtgagggcag aggaagtctg
     2221 ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgc tgctagaagc agaactcgat
     2281 tgccaccgag agaggcccgg tgcccctgga gcttctgccc tctgtacctt cagcaggact
     2341 ccagagatcc cgatgtgtgc aggctgtgac cagcacatct tggaccgttt catccttaag
     2401 gctctggacc gacattggca cagcaagtgt ctcaagtgca gtgactgcca cgtccctctg
     2461 gctgagcgct gcttcagccg cggggagagc gtctactgca aagacgactt ctttaagcgc
     2521 ttcgggacca agtgcgccgc atgccagctg ggcatcccgc ccacgcaggt ggtgcgccgc
     2581 gcccaggact tcgtgtacca cctgcattgc ttcgcctgcg tggtctgcaa gcggcagctg
     2641 gccacgggcg acgagttcta cctcatggaa gacagccggc tggtgtgcaa ggcggactac
     2701 gaaacagcca agcagcgaga agccgaggcc acagccaagc ggccgcgcac caccatcacc
     2761 gccaagcagc tggagacgct gaagagcgcc tacaacactt cgcccaagcc ggcgcgccac
     2821 gtgcgcgagc agctctcctc cgagactggc ctggacatgc gagtggtgca ggtgtggttc
     2881 cagaatcgcc gggctaagga aaagagactg aagaaagacg ctggccggca gcgctgggga
     2941 cagtatttcc gcaatatgaa gcgctcccgc ggcagttcca agtccgacaa ggacagcatc
     3001 caggagggac aagacagcga cgccgaagtc tccttcactg atgagccgtc catggctgac
     3061 atggggcctg ctaatggcct gtacagcagc ctgggagagc ctgcccctgc gttgggccgg
     3121 cccgtaggag gcctgggcag ctttaccctg gatcacggag gcttgacggg tccagagcag
     3181 taccgagagc tacgcccagg cagcccctat ggcatccccc catctcctgc agccccccag
     3241 agccttcctg gcccccagcc tctcctctcc agcctggtat acccagacac caacttgagc
     3301 cttgtccctt cagggccccc aggtggaccc ccacccatga gggtgctggc tggaaatggg
     3361 cccagctccg acctgtccac agagagcagt tctggctacc cagactttcc tgctagccct
     3421 gcttcctggc tggatgaagt agaccatgct cagttctaac gctgatatct agacccagct
     3481 ttcttgtaca aagtggttcg atagcttatc gataatcaac ctctggatta caaaatttgt
     3541 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3601 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3661 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3721 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3781 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3841 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3901 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3961 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4021 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4081 tccctttggg ccgcctcccc gcatcgatac cgtcgacgat aaaataaaag attttattta
     4141 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4201 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     4261 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     4321 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     4381 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     4441 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     4501 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     4561 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     4621 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     4681 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     4741 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     4801 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5461 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
     5761 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5821 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5881 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5941 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6001 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6061 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6121 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6181 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6241 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6301 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6361 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6421 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6481 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6541 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6601 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6661 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6721 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6781 gaaaagtgcc acctgcataa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     6841 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagaaaagt tcagatcaag
     6901 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     6961 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     7021 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     7081 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     7141 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     7201 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     7261 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     7321 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     7381 gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc
     7441 caccaccggg aggtaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct
     7501 atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg
     7561 acccgtggtg gaactgacga gttcggaaca cccggccgca accctgggag acgtcccagg
     7621 gacttcgggg gccgtttttg tggcccgacc tgagtccaaa aatcccgatc gttttggact
     7681 ctttggtgca cccccctaat aggagggata tgtggttctg gtaggagacg agaacctaaa
     7741 acagttcccg cctccgtctg aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg
     7801 tcttgtctgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     7861 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     7921 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     7981 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     8041 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     8101 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     8161 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     8221 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     8281 aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt
     8341 ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc
     8401 tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact
     8461 ggaccgaccg gtggtacctc acccttaccg agtcg
//
