LOCUS       sequence-463014-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Isl1 with mRuby2 fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  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-Isl1-P2A-mRuby2
FEATURES             Location/Qualifiers
     source          1..7154
                     /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"
     regulatory      1085..1094
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1351..1407
                     /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             1408..2118
                     /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(2135..2159)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2176..2764
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2229..2249)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2647..2658)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2944..3413
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3543..3560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3714..4302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3794..3813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4466..5257,5258..5326))
                     /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     5089..5108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5327..5431)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5461..6054
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6117..6474
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6533..6949
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6879..6901
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6917..6933
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6959..7154,1..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 caccatggga
      301 gacatgggcg atccaccaaa aaaaaaacgt ctgatttccc tgtgtgttgg ttgcggcaat
      361 caaattcacg accagtatat tctgagggtt tctccggatt tggagtggca tgcagcatgt
      421 ttgaaatgtg cggagtgtaa tcagtatttg gacgaaagct gtacgtgctt tgttagggat
      481 gggaaaacct actgtaaaag agattatatc aggttgtacg ggatcaaatg cgccaagtgc
      541 agcataggct tcagcaagaa cgacttcgtg atgcgcgccc gctctaaggt gtaccacatc
      601 gagtgtttcc gctgtgtagc ctgcagccga cagctcatcc cgggagacga attcgccctg
      661 cgggaggatg ggcttttctg ccgtgcagac cacgatgtgg tggagagagc cagcctggga
      721 gctggagatc ctctcagtcc cttgcatcca gcgcggcctc tgcaaatggc agccgaaccc
      781 atctcggcta ggcagccagc tctgcggccg cacgtccaca agcagccgga gaagaccacc
      841 cgagtgcgga ctgtgctcaa cgagaagcag ctgcacacct tgcggacctg ctatgccgcc
      901 aaccctcggc cagatgcgct catgaaggag caactagtgg agatgacggg cctcagtccc
      961 agagtcatcc gagtgtggtt tcaaaacaag cggtgcaagg acaagaaacg cagcatcatg
     1021 atgaagcagc tccagcagca gcaacccaac gacaaaacta atatccaggg gatgacagga
     1081 actcccatgg tggctgctag tccggagaga catgatggtg gtttacaggc taacccagta
     1141 gaggtgcaaa gttaccagcc gccctggaaa gtactgagtg acttcgcctt gcaaagcgac
     1201 atagatcagc ctgcttttca gcaactggtc aatttttcag aaggaggacc aggctctaat
     1261 tctactggca gtgaagtagc atcgatgtcc tcgcagctcc cagatacacc caacagcatg
     1321 gtagccagtc ctattgaggc aggaagcgga gctactaact tcagcctgct gaagcaggct
     1381 ggagacgtgg aggagaaccc tggacctatg gtgtctaagg gcgaagagct gatcaaggaa
     1441 aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca
     1501 ggtgaaggag aaggcaatcc gtacatggga actcaaacca tgaggatcaa agtcatcgag
     1561 ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt
     1621 acttttatca agtacccgaa aggcattcct gatttcttta aacagtcctt tcctgagggt
     1681 tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt catgcaggac
     1741 accagccttg aggatggctg tctcgtttac cacgtccaag tcagaggggt aaactttccc
     1801 tccaatggtc ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg
     1861 tatccagcag atggtggtct gaggggatac actcatatgg cactgaaagt tgatggtggt
     1921 ggccatctgt cttgctcttt cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc
     1981 aagatgcccg gtatccatgc cgttgatcac cgcctggaaa ggttagagga aagtgacaat
     2041 gaaatgttcg tagtacaacg cgaacacgca gttgccaagt tcgccgggct tggtggtggg
     2101 atggacgagc tgtacaagta acgctgatat ctagacccag ctttcttgta caaagtggtt
     2161 cgatagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2221 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2281 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2341 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2401 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2461 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2521 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2581 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2641 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2701 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2761 ccgcatcgat accgtcgacg ataaaataaa agattttatt tagtctccag aaaaaggggg
     2821 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     2881 catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa
     2941 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     3001 caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     3061 gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc
     3121 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     3181 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     3241 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     3301 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     3361 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag
     3421 catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg
     3481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4381 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg ccgcaaatca atctaaagta
     4441 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4501 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4561 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4621 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4681 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4741 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4801 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4861 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4921 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4981 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5041 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5101 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5161 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5221 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5281 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5341 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5401 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcat
     5461 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     5521 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
     5581 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     5641 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     5701 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     5761 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     5821 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     5881 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
     5941 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     6001 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     6061 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     6121 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
     6181 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     6241 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
     6301 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
     6361 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     6421 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     6481 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     6541 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     6601 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     6661 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     6721 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     6781 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     6841 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     6901 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     6961 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     7021 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     7081 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     7141 tcacccttac cgag
//
