LOCUS       sequence-463016-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of tagged mNgn2x3HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  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 6275)
  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-x3HA
FEATURES             Location/Qualifiers
     source          1..6275
                     /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"
     protein_bind    complement(1256..1280)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1297..1885
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1350..1370)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1768..1779)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2065..2534
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2664..2681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2835..3423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2915..2934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3587..4378,4379..4447))
                     /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     4210..4229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4448..4552)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4582..5175
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5238..5595
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5654..6070
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6000..6022
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6038..6054
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6080..6275,1..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 tggttctggt ggtgttgaat aacgctgata tctagaccca
     1261 gctttcttgt acaaagtggt tcgatagctt atcgataatc aacctctgga ttacaaaatt
     1321 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1381 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1441 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     1501 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     1561 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     1621 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     1681 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     1741 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     1801 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     1861 atctcccttt gggccgcctc cccgcatcga taccgtcgac gataaaataa aagattttat
     1921 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     1981 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     2041 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     2101 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     2161 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     2221 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     2281 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     2341 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     2401 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     2461 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     2521 gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt
     2581 tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg
     2641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3241 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg
     3541 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3601 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3661 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3721 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3781 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3841 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3901 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3961 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4021 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4081 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4141 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4201 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4261 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4321 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4381 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4441 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4501 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4561 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
     4621 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
     4681 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     4741 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     4801 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     4861 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     4921 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     4981 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     5041 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5101 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5161 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
     5221 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
     5281 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
     5341 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
     5401 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
     5461 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
     5521 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     5581 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     5641 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     5701 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     5761 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     5821 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     5881 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     5941 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     6001 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     6061 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     6121 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     6181 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     6241 actggaccga ccggtggtac ctcaccctta ccgag
//
