LOCUS       sequence-463001-        7520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mNgn2x3HA and Lhx3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7520)
  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 7520)
  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-mNgn2x3HA-T2A-mLhx3
FEATURES             Location/Qualifiers
     source          1..7520
                     /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"
     CDS             1123..1149
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1159..1185
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1195..1221
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1231..1284
                     /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(2501..2525)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2542..3130
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2595..2615)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3013..3024)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3310..3779
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3909..3926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4080..4668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4160..4179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4832..5623,5624..5692))
                     /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     5455..5474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5693..5797)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5827..6420
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6483..6840
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6899..7315
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7245..7267
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7283..7299
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(7325..7520,1..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 catgggaacc aattcagtcg actggatccg gtactactgc caccatgttc
      301 gtcaaatctg agactctgga gttgaaggag gaagaggagg tactgatgct gctgggctcg
      361 gcttccccgg cctcggcgac cctgaccccg atgtcctcca gcgcggacga ggaggaggac
      421 gaggagctgc gccggccggg ctccgcgcgt gggcagcgtg gagcggaagc cgggcagggg
      481 gtgcagggca gtccggcgtc gggtgccggg ggttgccggc cagggcggct gctgggcctg
      541 atgcacgagt gcaagcgtcg cccgtcgcgc tcacgggccg tctcccgagg tgccaagacg
      601 gcggagacgg tgcagcgcat caagaagacc cgcaggctca aggccaacaa ccgcgagcgc
      661 aaccgcatgc acaacctaaa cgccgcgctg gacgcgctgc gcgaggtgct gcccaccttc
      721 cccgaggatg ccaagctcac gaagatcgag acgctgcgct tcgcccacaa ttacatctgg
      781 gcgctcaccg agactctgcg cctggcggac cactgcgccg gcgccggtgg cctccagggg
      841 gcgctcttca cggaggcggt gctcctgagc ccgggagctg cgctcggcgc cagcggggac
      901 agcccttctc caccttcctc ctggagctgc accaacagcc cggcgtcatc ctccaactcc
      961 acgtccccat acagctgcac tttatcgccc gctagccccg ggtcagacgt ggactactgg
     1021 cagcccccac ctccggagaa gcatcgttat gcgcctcacc tgcccctcgc cagggactgt
     1081 atcgggtctg gctccggatc aagtggtggc agcggatcag gatatccata cgatgtacca
     1141 gactatgcgg gatcgggtta cccctacgac gtccccgact acgccggctc cggctacccc
     1201 tacgacgtcc ctgattatgc tggcagtggt gagggcagag gaagtctgct aacatgcggt
     1261 gacgtcgagg agaatcctgg cccaatgctg ctagaagcag aactcgattg ccaccgagag
     1321 aggcccggtg cccctggagc ttctgccctc tgtaccttca gcaggactcc agagatcccg
     1381 atgtgtgcag gctgtgacca gcacatcttg gaccgtttca tccttaaggc tctggaccga
     1441 cattggcaca gcaagtgtct caagtgcagt gactgccacg tccctctggc tgagcgctgc
     1501 ttcagccgcg gggagagcgt ctactgcaaa gacgacttct ttaagcgctt cgggaccaag
     1561 tgcgccgcat gccagctggg catcccgccc acgcaggtgg tgcgccgcgc ccaggacttc
     1621 gtgtaccacc tgcattgctt cgcctgcgtg gtctgcaagc ggcagctggc cacgggcgac
     1681 gagttctacc tcatggaaga cagccggctg gtgtgcaagg cggactacga aacagccaag
     1741 cagcgagaag ccgaggccac agccaagcgg ccgcgcacca ccatcaccgc caagcagctg
     1801 gagacgctga agagcgccta caacacttcg cccaagccgg cgcgccacgt gcgcgagcag
     1861 ctctcctccg agactggcct ggacatgcga gtggtgcagg tgtggttcca gaatcgccgg
     1921 gctaaggaaa agagactgaa gaaagacgct ggccggcagc gctggggaca gtatttccgc
     1981 aatatgaagc gctcccgcgg cagttccaag tccgacaagg acagcatcca ggagggacaa
     2041 gacagcgacg ccgaagtctc cttcactgat gagccgtcca tggctgacat ggggcctgct
     2101 aatggcctgt acagcagcct gggagagcct gcccctgcgt tgggccggcc cgtaggaggc
     2161 ctgggcagct ttaccctgga tcacggaggc ttgacgggtc cagagcagta ccgagagcta
     2221 cgcccaggca gcccctatgg catcccccca tctcctgcag ccccccagag ccttcctggc
     2281 ccccagcctc tcctctccag cctggtatac ccagacacca acttgagcct tgtcccttca
     2341 gggcccccag gtggaccccc acccatgagg gtgctggctg gaaatgggcc cagctccgac
     2401 ctgtccacag agagcagttc tggctaccca gactttcctg ctagccctgc ttcctggctg
     2461 gatgaagtag accatgctca gttctaacgc tgatatctag acccagcttt cttgtacaaa
     2521 gtggttcgat agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2581 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2641 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2701 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2761 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2821 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2881 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     2941 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3001 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3061 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3121 gcctccccgc atcgataccg tcgacgataa aataaaagat tttatttagt ctccagaaaa
     3181 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     3241 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     3301 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3361 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     3421 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     3481 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     3541 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3601 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     3661 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     3721 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     3781 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     3841 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3901 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3961 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4021 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4081 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4141 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4201 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4261 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4321 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4381 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4441 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4501 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4561 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4621 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4681 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4741 atgagattat caaaaaggat cttcacctag atccttttgc ggccggccgc aaatcaatct
     4801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5821 ctgcataatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     5881 caaggcatgg aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaga
     5941 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6001 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     6061 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     6121 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     6181 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     6241 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga ttgactgagt
     6301 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     6361 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     6421 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
     6481 gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
     6541 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
     6601 actgacgagt tcggaacacc cggccgcaac cctgggagac gtcccaggga cttcgggggc
     6661 cgtttttgtg gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc
     6721 cccctaatag gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     6781 tccgtctgaa tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg
     6841 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     6901 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     6961 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     7021 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     7081 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     7141 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     7201 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     7261 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     7321 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     7381 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     7441 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     7501 ggtacctcac ccttaccgag
//
