LOCUS       sequence-488749-        7241 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  human PAX3::FOXO1 with homeobox domain deletion in -2A-sfGFP 
            backbone (Plasmid# 240099).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7241)
  AUTHORS   Kucinski J, Tallan A, Taslim C, Vontell AM, Silvius KM, Wang M, 
            Cannon MV, Stanton BZ, Kendall GC
  TITLE     Rhabdomyosarcoma fusion oncoprotein initially pioneers a neural 
            signature in vivo.
  JOURNAL   Cell Rep. 2025 Jul 22;44(7):115923. doi: 
            10.1016/j.celrep.2025.115923. Epub 2025 Jul 3.
  PUBMED    40614256
REFERENCE   2  (bases 1 to 7241)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2025-07-22"; volume: "44(7):115923. doi"; pages: " 
            10.1016/j.celrep.2025.115923. Epub 2025 Jul 3"
COMMENT     Sequence Label: PAX3::FOXO1-Delta-HD-2A-sfGFP
FEATURES             Location/Qualifiers
     source          1..7241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2484..2540
                     /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             join(2547..2549,2550..3263)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2547..3260
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2577..2605)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3200..3220
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3293..3427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3317..3336)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3371..3390
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3540..3560)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3540..3558)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3579..3595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3579..3595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3592..3614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3603..3619
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3627..3633,3634..3651,3652..3657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3672..3693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3981..4569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4061..4080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4740..5531,5532..5600))
                     /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     5363..5382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5601..5705)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5731..6186)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5868..5889)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6080..6099
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6257..7241
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggcctctcg agcctctaga
      121 ttaattaagc cgccaccatg accacgctgg ccggcgctgt gcccaggatg atgcggccgg
      181 gcccggggca gaactacccg cgtagcgggt tcccgctgga agtgtccact cccctcggcc
      241 agggccgcgt caaccagctc ggcggcgttt ttatcaacgg caggccgctg cccaaccaca
      301 tccgccacaa gatcgtggag atggcccacc acggcatccg gccctgcgtc atctcgcgcc
      361 agctgcgcgt gtcccacggc tgcgtctcca agatcctgtg caggtaccag gagactggct
      421 ccatacgtcc tggtgccatc ggcggcagca agcccaagca ggtgacaacg cctgacgtgg
      481 agaagaaaat tgaggaatac aaaagagaga acccgggcat gttcagctgg gaaatccgag
      541 acaaattact caaggacgcg gtctgtgatc gaaacaccgt gccgtcagtg agttccatca
      601 gccgcatcct gagaagtaaa ttcgggaaag gtgaagagga ggaggccgac ttggagagga
      661 aggaggcaga ggaaagcgag aagaaggcca aacacagcat cgacggcatc ctgagcgagc
      721 gagcctcagc accccaatca gatgaaggct ctgatattga ctctgaacca gatttaccac
      781 taaagaggaa aggggccaat caactgatgg ctttcaacca tctcattccc ggggggttcc
      841 ctcccactgc catgccgacc ttgccaacgt accagctgtc ggagacctct taccagccca
      901 catctattcc acaagctgtg tcagatccca gcagcaccgt tcacagacct caaccgcttc
      961 ctccaagcac tgtacaccaa agcacgattc cttccaaccc agacagcagc tctgcctact
     1021 gcctccccag caccaggcat ggattttcca gctatacaga cagctttgtg cctccgtcgg
     1081 ggccctccaa ccccatgaac cccaccattg gcaatggcct ctcacctcag aattcaattc
     1141 gtcataatct gtccctacac agcaagttca ttcgtgtgca gaatgaagga actggaaaaa
     1201 gttcttggtg gatgctcaat ccagagggtg gcaagagcgg gaaatctcct aggagaagag
     1261 ctgcatccat ggacaacaac agtaaatttg ctaagagccg aagccgagct gccaagaaga
     1321 aagcatctct ccagtctggc caggagggtg ctggggacag ccctggatca cagttttcca
     1381 aatggcctgc aagccctggc tctcacagca atgatgactt tgataactgg agtacatttc
     1441 gccctcgaac tagctcaaat gctagtacta ttagtgggag actctcaccc attatgaccg
     1501 aacaggatga tcttggagaa ggggatgtgc attctatggt gtacccgcca tctgccgcaa
     1561 agatggcctc tactttaccc agtctgtctg agataagcaa tcccgaaaac atggaaaatc
     1621 ttttggataa tctcaacctt ctctcatcac caacatcatt aactgtttcg acccagtcct
     1681 cacctggcac catgatgcag cagacgccgt gctactcgtt tgcgccacca aacaccagtt
     1741 tgaattcacc cagcccaaac taccaaaaat atacatatgg ccaatccagc atgagccctt
     1801 tgccccagat gcctatacaa acacttcagg acaataagtc gagttatgga ggtatgagtc
     1861 agtataactg tgcgcctgga ctcttgaagg agttgctgac ttctgactct cctccccata
     1921 atgacattat gacaccagtt gatcctgggg tagcccagcc caacagccgg gttctgggcc
     1981 agaacgtcat gatgggccct aattcggtca tgtcaaccta tggcagccag gcatctcata
     2041 acaaaatgat gaatcccagc tcccataccc accctggaca tgctcagcag acatctgcag
     2101 ttaacgggcg tcccctgccc cacacggtaa gcaccatgcc ccacacctcg ggtatgaacc
     2161 gcctgaccca agtgaagaca cctgtacaag tgcctctgcc ccaccccatg cagatgagtg
     2221 ccctgggggg ctactcctcc gtgagcagct gcaatggcta tggcagaatg ggccttctcc
     2281 accaggagaa gctcccaagt gacttggatg gcatgttcat tgagcgctta gactgtgaca
     2341 tggaatccat cattcggaat gacctcatgg atggagatac attggatttt aactttgaca
     2401 atgtgttgcc caaccaaagc ttcccacaca gtgtcaagac aacgacacat agctgggtgt
     2461 caggcggcgc gccaggaagc ggagctacta acttcagcct gctgaagcag gctggagacg
     2521 tggaggagaa ccctggacct ggatccatga gcaaaggaga agaacttttc actggagttg
     2581 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtccgtggag
     2641 agggtgaagg tgatgctaca aacggaaaac tcacccttaa atttatttgc actactggaa
     2701 aactacctgt tccgtggcca acacttgtca ctactctgac ctatggtgtt caatgctttt
     2761 cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt
     2821 atgtacagga acgcactata tctttcaaag atgacgggac ctacaagacg cgtgctgaag
     2881 tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag
     2941 aagatggaaa cattcttgga cacaaactcg agtacaactt taactcacac aatgtataca
     3001 tcacggcaga caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacgttg
     3061 aagatggttc cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     3121 ctgtcctttt accagacaac cattacctgt cgacacaatc tgtcctgtcg aaagatccca
     3181 acgaaaagcg tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg
     3241 gcatggatga gctctacaaa taatctagaa ctatagtgag tcgtattacg tagatccaga
     3301 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3361 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3421 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3481 ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc
     3541 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     3601 aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc
     3661 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3721 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3781 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5701 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     5761 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5821 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5881 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5941 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6001 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6061 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     6121 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     6181 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6241 cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt
     6301 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
     6361 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
     6421 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
     6481 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
     6541 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
     6601 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
     6661 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
     6721 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
     6781 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
     6841 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
     6901 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
     6961 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
     7021 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
     7081 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     7141 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     7201 cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
