LOCUS       sequence-463022-        6064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Ngn2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  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 6064)
  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-mNgn2
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    397..754
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             813..1229
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1159..1181
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1197..1213
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1239..1613
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1551..1567
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    1656..1680
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2479..2503)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2520..3108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2573..2593)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2991..3002)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3288..3757
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3887..3904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4058..4646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4138..4157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4810..5601,5602..5670))
                     /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     5433..5452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5671..5775)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(5805..6064,1..334)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
       61 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
      121 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
      181 tcactcggcg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
      241 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
      301 gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac
      361 ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg
      421 tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt actagttagc
      481 taactagctc tgtatctggc ggacccgtgg tggaactgac gagttcggaa cacccggccg
      541 caaccctggg agacgtccca gggacttcgg gggccgtttt tgtggcccga cctgagtcca
      601 aaaatcccga tcgttttgga ctctttggtg caccccccta ataggaggga tatgtggttc
      661 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
      721 gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc
      781 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
      841 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
      901 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
      961 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1021 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1081 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     1141 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     1201 atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg
     1261 cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct
     1321 ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg
     1381 cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg
     1441 acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt
     1501 acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac
     1561 acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctaga ctgccggatc
     1621 tagctagtta atcgacggta tcgataagct atcaaacaag tttgtacaaa aaagcaggct
     1681 gccaccatgt tcgtcaaatc tgagactctg gagttgaagg aggaagagga ggtactgatg
     1741 ctgctgggct cggcttcccc ggcctcggcg accctgaccc cgatgtcctc cagcgcggac
     1801 gaggaggagg acgaggagct gcgccggccg ggctccgcgc gtgggcagcg tggagcggaa
     1861 gccgggcagg gggtgcaggg cagtccggcg tcgggtgccg ggggttgccg gccagggcgg
     1921 ctgctgggcc tgatgcacga gtgcaagcgt cgcccgtcgc gctcacgggc cgtctcccga
     1981 ggtgccaaga cggcggagac ggtgcagcgc atcaagaaga cccgcaggct caaggccaac
     2041 aaccgcgagc gcaaccgcat gcacaaccta aacgccgcgc tggacgcgct gcgcgaggtg
     2101 ctgcccacct tccccgagga tgccaagctc acgaagatcg agacgctgcg cttcgcccac
     2161 aattacatct gggcgctcac cgagactctg cgcctggcgg accactgcgc cggcgccggt
     2221 ggcctccagg gggcgctctt cacggaggcg gtgctcctga gcccgggagc tgcgctcggc
     2281 gccagcgggg acagcccttc tccaccttcc tcctggagct gcaccaacag cccggcgtca
     2341 tcctccaact ccacgtcccc atacagctgc actttatcgc ccgctagccc cgggtcagac
     2401 gtggactact ggcagccccc acctccggag aagcatcgtt atgcgcctca cctgcccctc
     2461 gccagggact gtatctagac ccagctttct tgtacaaagt ggttcgatag cttatcgata
     2521 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2581 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2641 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2701 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2761 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2821 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2881 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2941 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3001 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3061 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3121 gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     3181 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     3241 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     3301 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     3361 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3421 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     3481 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3541 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3601 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3661 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3721 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     3781 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     3841 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4741 tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg
     4801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcataatgaa agaccccacc
     5821 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     5881 ctgagaatag aaaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     5941 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     6001 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     6061 agaa
//
