LOCUS       sequence-463006-        7307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mLhx3 with mRuby2 fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7307)
  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 7307)
  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-mLhx3-P2A-mRuby2
FEATURES             Location/Qualifiers
     source          1..7307
                     /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             1504..1560
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1561..2271
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     protein_bind    complement(2288..2312)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2329..2917
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2382..2402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2800..2811)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3097..3566
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3696..3713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3867..4455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3947..3966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4619..5410,5411..5479))
                     /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     5242..5261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5480..5584)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5614..6207
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6270..6627
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6686..7102
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7032..7054
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7070..7086
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(7112..7307,1..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 caccatgctg
      301 ctagaagcag aactcgattg ccaccgagag aggcccggtg cccctggagc ttctgccctc
      361 tgtaccttca gcaggactcc agagatcccg atgtgtgcag gctgtgacca gcacatcttg
      421 gaccgtttca tccttaaggc tctggaccga cattggcaca gcaagtgtct caagtgcagt
      481 gactgccacg tccctctggc tgagcgctgc ttcagccgcg gggagagcgt ctactgcaaa
      541 gacgacttct ttaagcgctt cgggaccaag tgcgccgcat gccagctggg catcccgccc
      601 acgcaggtgg tgcgccgcgc ccaggacttc gtgtaccacc tgcattgctt cgcctgcgtg
      661 gtctgcaagc ggcagctggc cacgggcgac gagttctacc tcatggaaga cagccggctg
      721 gtgtgcaagg cggactacga aacagccaag cagcgagaag ccgaggccac agccaagcgg
      781 ccgcgcacca ccatcaccgc caagcagctg gagacgctga agagcgccta caacacttcg
      841 cccaagccgg cgcgccacgt gcgcgagcag ctctcctccg agactggcct ggacatgcga
      901 gtggtgcagg tgtggttcca gaatcgccgg gctaaggaaa agagactgaa gaaagacgct
      961 ggccggcagc gctggggaca gtatttccgc aatatgaagc gctcccgcgg cagttccaag
     1021 tccgacaagg acagcatcca ggagggacaa gacagcgacg ccgaagtctc cttcactgat
     1081 gagccgtcca tggctgacat ggggcctgct aatggcctgt acagcagcct gggagagcct
     1141 gcccctgcgt tgggccggcc cgtaggaggc ctgggcagct ttaccctgga tcacggaggc
     1201 ttgacgggtc cagagcagta ccgagagcta cgcccaggca gcccctatgg catcccccca
     1261 tctcctgcag ccccccagag ccttcctggc ccccagcctc tcctctccag cctggtatac
     1321 ccagacacca acttgagcct tgtcccttca gggcccccag gtggaccccc acccatgagg
     1381 gtgctggctg gaaatgggcc cagctccgac ctgtccacag agagcagttc tggctaccca
     1441 gactttcctg ctagccctgc ttcctggctg gatgaagtag accatgctca gttcggaagc
     1501 ggagctacta acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct
     1561 atggtgtcta agggcgaaga gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa
     1621 ggttcggtca acggccacca attcaaatgc acaggtgaag gagaaggcaa tccgtacatg
     1681 ggaactcaaa ccatgaggat caaagtcatc gagggaggac ccctgccatt tgcctttgac
     1741 attcttgcca cgtcgttcat gtatggcagc cgtactttta tcaagtaccc gaaaggcatt
     1801 cctgatttct ttaaacagtc ctttcctgag ggttttactt gggaaagagt tacgagatac
     1861 gaagatggtg gagtcgtcac cgtcatgcag gacaccagcc ttgaggatgg ctgtctcgtt
     1921 taccacgtcc aagtcagagg ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag
     1981 accaagggtt gggagcctaa tacagagatg atgtatccag cagatggtgg tctgagggga
     2041 tacactcata tggcactgaa agttgatggt ggtggccatc tgtcttgctc tttcgtaaca
     2101 acttacaggt caaaaaagac cgtcgggaac atcaagatgc ccggtatcca tgccgttgat
     2161 caccgcctgg aaaggttaga ggaaagtgac aatgaaatgt tcgtagtaca acgcgaacac
     2221 gcagttgcca agttcgccgg gcttggtggt gggatggacg agctgtacaa gtaacgctga
     2281 tatctagacc cagctttctt gtacaaagtg gttcgatagc ttatcgataa tcaacctctg
     2341 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2401 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2461 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2521 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2581 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2641 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2701 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     2761 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2821 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2881 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acgataaaat
     2941 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     3001 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     3061 agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
     3121 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     3181 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3241 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     3301 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3361 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3421 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     3481 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     3541 gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt
     3601 ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg
     3661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4561 cttttgcggc cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4741 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5581 cgcgcacatt tccccgaaaa gtgccacctg cataatgaaa gaccccacct gtaggtttgg
     5641 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     5701 aaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     5761 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     5821 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     5881 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     5941 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     6001 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     6061 gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     6121 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     6181 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     6241 cagggaccac cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc
     6301 gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag
     6361 ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg ccgcaaccct
     6421 gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt ccaaaaatcc
     6481 cgatcgtttt ggactctttg gtgcaccccc ctaataggag ggatatgtgg ttctggtagg
     6541 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttgggaccg
     6601 aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct gtctgactgt
     6661 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     6721 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     6781 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     6841 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     6901 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     6961 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     7021 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     7081 cctcactcct tctctaggcg cccccatatg gccatatgag atcttatatg gggcaccccc
     7141 gccccttgta aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca
     7201 agctcactta caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc
     7261 ctaccaagaa caactggacc gaccggtggt acctcaccct taccgag
//
