LOCUS       sequence-462999-        6062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of fluorescent mRuby2 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  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 6062)
  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-mRuby2
FEATURES             Location/Qualifiers
     source          1..6062
                     /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"
     regulatory      289..298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             295..1005
                     /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(1043..1067)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1084..1672
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1137..1157)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1555..1566)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1852..2321
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2451..2468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2622..3210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2702..2721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3374..4165,4166..4234))
                     /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     3997..4016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4235..4339)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4369..4962
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5025..5382
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5441..5857
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5787..5809
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5825..5841
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(5867..6062,1..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 gtacaatggc caccatggtg
      301 tctaagggcg aagagctgat caaggaaaat atgcgtatga aggtggtcat ggaaggttcg
      361 gtcaacggcc accaattcaa atgcacaggt gaaggagaag gcaatccgta catgggaact
      421 caaaccatga ggatcaaagt catcgaggga ggacccctgc catttgcctt tgacattctt
      481 gccacgtcgt tcatgtatgg cagccgtact tttatcaagt acccgaaagg cattcctgat
      541 ttctttaaac agtcctttcc tgagggtttt acttgggaaa gagttacgag atacgaagat
      601 ggtggagtcg tcaccgtcat gcaggacacc agccttgagg atggctgtct cgtttaccac
      661 gtccaagtca gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa gaagaccaag
      721 ggttgggagc ctaatacaga gatgatgtat ccagcagatg gtggtctgag gggatacact
      781 catatggcac tgaaagttga tggtggtggc catctgtctt gctctttcgt aacaacttac
      841 aggtcaaaaa agaccgtcgg gaacatcaag atgcccggta tccatgccgt tgatcaccgc
      901 ctggaaaggt tagaggaaag tgacaatgaa atgttcgtag tacaacgcga acacgcagtt
      961 gccaagttcg ccgggcttgg tggtgggatg gacgagctgt acaagtaaca acaagcttga
     1021 taccctagga ggagatatct agacccagct ttcttgtaca aagtggttcg atagcttatc
     1081 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1141 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1201 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1261 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1321 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     1381 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1441 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     1501 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     1561 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     1621 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     1681 cgtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     1741 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     1801 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     1861 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     1921 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     1981 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     2041 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     2101 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     2161 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     2221 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     2281 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     2341 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     2401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3301 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     3361 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3601 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3661 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3781 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3901 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4021 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4081 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4201 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4261 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4321 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcataa tgaaagaccc
     4381 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     4441 ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     4501 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4561 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4621 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     4681 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     4741 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     4801 acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     4861 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     4921 tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat
     4981 cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact
     5041 tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac
     5101 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttcggaaca
     5161 cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg tggcccgacc
     5221 tgagtccaaa aatcccgatc gttttggact ctttggtgca cccccctaat aggagggata
     5281 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
     5341 ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg ttctgtgttg
     5401 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     5461 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     5521 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     5581 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     5641 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     5701 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     5761 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     5821 ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat atgagatctt
     5881 atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa gagttactaa
     5941 cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg aagtctggag
     6001 acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc acccttaccg
     6061 ag
//
