LOCUS       sequence-462995-        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of tagged mNgn2x3FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Wang NB, Adewumi HO, Lende-Dorn BA, Beitz AM, O'Shea TM, Galloway KE
  TITLE     Compact transcription factor cassettes generate functional, 
            engraftable motor neurons by direct conversion.
  JOURNAL   Cell Syst. 2025 Mar 10:101206. doi: 10.1016/j.cels.2025.101206.
  PUBMED    40086435
REFERENCE   2  (bases 1 to 6194)
  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. 
            2025 Mar 10:101206. doi: 10.1016/j.cels.2025.101206."
COMMENT     Sequence Label: pMXs-mNgn2-x3FLAG
FEATURES             Location/Qualifiers
     source          1..6194
                     /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             1093..1158
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1175..1199)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1216..1804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1269..1289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1687..1698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1984..2453
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2583..2600)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2754..3342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2834..2853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3506..4297,4298..4366))
                     /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     4129..4148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4367..4471)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4501..5094
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5157..5514
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5573..5989
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5919..5941
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5957..5973
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(5999..6194,1..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 atcggaagcg gtgattataa ggatcacgat ggcgactaca aggaccacga tattgactac
     1141 aaagacgatg acgataaatg acgctgatat ctagacccag ctttcttgta caaagtggtt
     1201 cgatagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1261 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1321 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     1381 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     1441 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     1501 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1561 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     1621 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     1681 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     1741 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     1801 ccgcatcgat accgtcgacg ataaaataaa agattttatt tagtctccag aaaaaggggg
     1861 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     1921 catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa
     1981 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     2041 caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     2101 gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc
     2161 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     2221 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     2281 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     2341 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     2401 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag
     2461 catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg
     2521 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2581 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2641 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3181 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3421 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg ccgcaaatca atctaaagta
     3481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcat
     4501 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     4561 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
     4621 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4681 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     4741 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     4801 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     4861 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     4921 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
     4981 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5041 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     5101 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     5161 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
     5221 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     5281 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
     5341 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
     5401 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     5461 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     5521 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     5581 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     5641 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     5701 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     5761 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     5821 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     5881 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     5941 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     6001 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     6061 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     6121 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     6181 tcacccttac cgag
//
