LOCUS       sequence-463017-        7630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Isl1 and Lhx3 for motor neuron 
            specification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7630)
  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 7630)
  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-Isl1-T2A-mLhx3
FEATURES             Location/Qualifiers
     source          1..7630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..133
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    222..246
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      283..292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1345..1398
                     /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(2611..2635)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2652..3240
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2705..2725)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3123..3134)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3420..3889
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4019..4036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4942..5733,5734..5802))
                     /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     5565..5584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5803..5907)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5937..6530
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6593..6950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7009..7425
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7355..7377
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7393..7409
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(7435..7630,1..179)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag
       61 gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt
      121 ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc
      181 cggatctagc tagttaatcg acggtatcga taagctatca aacaagtttg tacaaaaaag
      241 caggctccac caggaaccaa ttcagtcgac tggatccggt acgccaccat gggagacatg
      301 ggcgatccac caaaaaaaaa acgtctgatt tccctgtgtg ttggttgcgg caatcaaatt
      361 cacgaccagt atattctgag ggtttctccg gatttggagt ggcatgcagc atgtttgaaa
      421 tgtgcggagt gtaatcagta tttggacgaa agctgtacgt gctttgttag ggatgggaaa
      481 acctactgta aaagagatta tatcaggttg tacgggatca aatgcgccaa gtgcagcata
      541 ggcttcagca agaacgactt cgtgatgcgc gcccgctcta aggtgtacca catcgagtgt
      601 ttccgctgtg tagcctgcag ccgacagctc atcccgggag acgaattcgc cctgcgggag
      661 gatgggcttt tctgccgtgc agaccacgat gtggtggaga gagccagcct gggagctgga
      721 gatcctctca gtcccttgca tccagcgcgg cctctgcaaa tggcagccga acccatctcg
      781 gctaggcagc cagctctgcg gccgcacgtc cacaagcagc cggagaagac cacccgagtg
      841 cggactgtgc tcaacgagaa gcagctgcac accttgcgga cctgctatgc cgccaaccct
      901 cggccagatg cgctcatgaa ggagcaacta gtggagatga cgggcctcag tcccagagtc
      961 atccgagtgt ggtttcaaaa caagcggtgc aaggacaaga aacgcagcat catgatgaag
     1021 cagctccagc agcagcaacc caacgacaaa actaatatcc aggggatgac aggaactccc
     1081 atggtggctg ctagtccgga gagacatgat ggtggtttac aggctaaccc agtagaggtg
     1141 caaagttacc agccgccctg gaaagtactg agtgacttcg ccttgcaaag cgacatagat
     1201 cagcctgctt ttcagcaact ggtcaatttt tcagaaggag gaccaggctc taattctact
     1261 ggcagtgaag tagcatcgat gtcctcgcag ctcccagata cacccaacag catggtagcc
     1321 agtcctattg aggcaggcag tggtgagggc agaggaagtc tgctaacatg cggtgacgtc
     1381 gaggagaatc ctggcccaat gctgctagaa gcagaactcg attgccaccg agagaggccc
     1441 ggtgcccctg gagcttctgc cctctgtacc ttcagcagga ctccagagat cccgatgtgt
     1501 gcaggctgtg accagcacat cttggaccgt ttcatcctta aggctctgga ccgacattgg
     1561 cacagcaagt gtctcaagtg cagtgactgc cacgtccctc tggctgagcg ctgcttcagc
     1621 cgcggggaga gcgtctactg caaagacgac ttctttaagc gcttcgggac caagtgcgcc
     1681 gcatgccagc tgggcatccc gcccacgcag gtggtgcgcc gcgcccagga cttcgtgtac
     1741 cacctgcatt gcttcgcctg cgtggtctgc aagcggcagc tggccacggg cgacgagttc
     1801 tacctcatgg aagacagccg gctggtgtgc aaggcggact acgaaacagc caagcagcga
     1861 gaagccgagg ccacagccaa gcggccgcgc accaccatca ccgccaagca gctggagacg
     1921 ctgaagagcg cctacaacac ttcgcccaag ccggcgcgcc acgtgcgcga gcagctctcc
     1981 tccgagactg gcctggacat gcgagtggtg caggtgtggt tccagaatcg ccgggctaag
     2041 gaaaagagac tgaagaaaga cgctggccgg cagcgctggg gacagtattt ccgcaatatg
     2101 aagcgctccc gcggcagttc caagtccgac aaggacagca tccaggaggg acaagacagc
     2161 gacgccgaag tctccttcac tgatgagccg tccatggctg acatggggcc tgctaatggc
     2221 ctgtacagca gcctgggaga gcctgcccct gcgttgggcc ggcccgtagg aggcctgggc
     2281 agctttaccc tggatcacgg aggcttgacg ggtccagagc agtaccgaga gctacgccca
     2341 ggcagcccct atggcatccc cccatctcct gcagcccccc agagccttcc tggcccccag
     2401 cctctcctct ccagcctggt atacccagac accaacttga gccttgtccc ttcagggccc
     2461 ccaggtggac ccccacccat gagggtgctg gctggaaatg ggcccagctc cgacctgtcc
     2521 acagagagca gttctggcta cccagacttt cctgctagcc ctgcttcctg gctggatgaa
     2581 gtagaccatg ctcagttcta agatatctag acccagcttt cttgtacaaa gtggttcgat
     2641 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2701 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2761 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2821 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2881 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2941 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3001 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3061 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3121 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3181 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3241 atcgataccg tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3301 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3361 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     3421 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3481 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3541 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     3601 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3661 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3721 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     3781 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     3841 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     3901 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     3961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata
     4921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4981 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5041 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5101 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5161 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5221 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5281 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5341 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5401 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5461 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5521 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5581 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5641 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5701 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5761 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5881 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcataatg
     5941 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     6001 aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacagct
     6061 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6121 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     6181 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
     6241 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     6301 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     6361 aagagcccac aacccctcac tcggcgcgcc agtcctccga ttgactgagt cgcccgggta
     6421 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     6481 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
     6541 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
     6601 cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc
     6661 gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     6721 tcggaacacc cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg
     6781 gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc cccctaatag
     6841 gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     6901 tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt
     6961 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
     7021 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     7081 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     7141 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     7201 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     7261 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     7321 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     7381 cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat atggccatat
     7441 gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga catgacaaga
     7501 gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt ccagcacgaa
     7561 gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt ggtacctcac
     7621 ccttaccgag
//
