LOCUS       sequence-471078-        7367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mNgn2x3HA and Isl1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7367)
  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 7367)
  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-Isl1
FEATURES             Location/Qualifiers
     source          1..7367
                     /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"
     regulatory      2075..2084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2348..2372)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2389..2977
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2442..2462)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2860..2871)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3157..3626
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3756..3773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3927..4515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4007..4026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4679..5470,5471..5539))
                     /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     5302..5321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5540..5644)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5674..6267
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6330..6687
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6746..7162
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7092..7114
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7130..7146
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(7172..7367,1..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 cccaatggga gacatgggcg atccaccaaa aaaaaaacgt
     1321 ctgatttccc tgtgtgttgg ttgcggcaat caaattcacg accagtatat tctgagggtt
     1381 tctccggatt tggagtggca tgcagcatgt ttgaaatgtg cggagtgtaa tcagtatttg
     1441 gacgaaagct gtacgtgctt tgttagggat gggaaaacct actgtaaaag agattatatc
     1501 aggttgtacg ggatcaaatg cgccaagtgc agcataggct tcagcaagaa cgacttcgtg
     1561 atgcgcgccc gctctaaggt gtaccacatc gagtgtttcc gctgtgtagc ctgcagccga
     1621 cagctcatcc cgggagacga attcgccctg cgggaggatg ggcttttctg ccgtgcagac
     1681 cacgatgtgg tggagagagc cagcctggga gctggagatc ctctcagtcc cttgcatcca
     1741 gcgcggcctc tgcaaatggc agccgaaccc atctcggcta ggcagccagc tctgcggccg
     1801 cacgtccaca agcagccgga gaagaccacc cgagtgcgga ctgtgctcaa cgagaagcag
     1861 ctgcacacct tgcggacctg ctatgccgcc aaccctcggc cagatgcgct catgaaggag
     1921 caactagtgg agatgacggg cctcagtccc agagtcatcc gagtgtggtt tcaaaacaag
     1981 cggtgcaagg acaagaaacg cagcatcatg atgaagcagc tccagcagca gcaacccaac
     2041 gacaaaacta atatccaggg gatgacagga actcccatgg tggctgctag tccggagaga
     2101 catgatggtg gtttacaggc taacccagta gaggtgcaaa gttaccagcc gccctggaaa
     2161 gtactgagtg acttcgcctt gcaaagcgac atagatcagc ctgcttttca gcaactggtc
     2221 aatttttcag aaggaggacc aggctctaat tctactggca gtgaagtagc atcgatgtcc
     2281 tcgcagctcc cagatacacc caacagcatg gtagccagtc ctattgaggc atgacgctga
     2341 tatctagacc cagctttctt gtacaaagtg gttcgatagc ttatcgataa tcaacctctg
     2401 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2461 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2521 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2581 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2641 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2701 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2761 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     2821 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2881 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2941 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acgataaaat
     3001 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     3061 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     3121 agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
     3181 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     3241 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3301 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     3361 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3421 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3481 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     3541 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     3601 gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt
     3661 ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg
     3721 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3901 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4081 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4201 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4261 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4321 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4381 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4441 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4501 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4561 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4621 cttttgcggc cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4981 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5281 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5641 cgcgcacatt tccccgaaaa gtgccacctg cataatgaaa gaccccacct gtaggtttgg
     5701 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     5761 aaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     5821 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     5881 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     5941 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     6001 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     6061 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     6121 gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     6181 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     6241 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     6301 cagggaccac cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc
     6361 gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag
     6421 ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg ccgcaaccct
     6481 gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt ccaaaaatcc
     6541 cgatcgtttt ggactctttg gtgcaccccc ctaataggag ggatatgtgg ttctggtagg
     6601 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttgggaccg
     6661 aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct gtctgactgt
     6721 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     6781 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     6841 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     6901 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     6961 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     7021 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     7081 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     7141 cctcactcct tctctaggcg cccccatatg gccatatgag atcttatatg gggcaccccc
     7201 gccccttgta aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca
     7261 agctcactta caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc
     7321 ctaccaagaa caactggacc gaccggtggt acctcaccct taccgag
//
