LOCUS       sequence-407515-        8571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, FUS_FLI1 from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8571)
  AUTHORS   Tripathi S, Shirnekhi HK, Gorman SD, Chandra B, Baggett DW, Park CG,
            Somjee R, Lang B, Hosseini SMH, Pioso BJ, Li Y, Iacobucci I, Gao Q, 
            Edmonson MN, Rice SV, Zhou X, Bollinger J, Mitrea DM, White MR, 
            McGrail DJ, Jarosz DF, Yi SS, Babu MM, Mullighan CG, Zhang J, Sahni 
            N, Kriwacki RW
  TITLE     Defining the condensate landscape of fusion oncoproteins.
  JOURNAL   Nat Commun. 2023 Sep 28;14(1):6008. doi: 10.1038/s41467-023-41655-2.
  PUBMED    37770423
REFERENCE   2  (bases 1 to 8571)
  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: "Nat 
            Commun."; date: "2023-09-28"; volume: "14(1):6008. doi"; pages: " 
            10.1038/s41467-023-41655-2"
COMMENT     Sequence Label: CL20_mEGFP_FUS_FLI1
FEATURES             Location/Qualifiers
     source          1..8571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    558..692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(582..601)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     636..655
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(698..893)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      712..847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    952..973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(988..993,994..1011,1012..1018)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1026..1042
                     /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)."
     primer_bind     1031..1053
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1050..1066
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1050..1066
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1409..1428
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1427..1482
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1528..1547
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3332..4042,4043..4045,4046..4048))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3374..3395)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3723..3742
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3982..4003
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4045..4054
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4130..4333)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4159..4179)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4334..4637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4894..4938)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5123..5356
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    5716..5833
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6325..6450
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6497..6754
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6756..7135
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7166..7270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7271..7339,7340..8131)
                     /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     complement(7489..7508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8302..8571,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca
      601 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
      661 ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa
      721 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
      781 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
      841 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg
      901 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      961 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1021 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1081 gcttgggctg caggtcgagg gatctccata agagaagagg gacagctatg actgggagta
     1141 gtcaggagag gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta
     1201 aagaaaaaaa taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc
     1261 tatggaataa ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc
     1321 aacctttgtt catggcagcc agcatatggg catatgttgc caaactctaa accaaatact
     1381 cattctgatg ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa
     1441 aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct
     1501 caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatct gctagctcga
     1561 gcagctgaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     1621 ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagat cttgtcttct
     1681 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
     1741 agctgccttg taagtcattg gtcttaaagg taccgtaatc ttatctagaa ccggttcagt
     1801 agtaagaccc gaggtgtgat ggcacatgag tgttagggtg tcgaggaaca ttaggattgg
     1861 gatagatccc acccgtcggg gatgtccagt attgggatgc cgcaccaaag aatgaacttg
     1921 aagttaccgg cattgagcta gggtgtggcg gtacgaaatt aaccttctgc tgatgcgcgt
     1981 gataggaggg catatagctg atgtcgctcg ggtatttata cattgagctc tcagtaggat
     2041 ggggttgcaa ggcttgtgca ataccgtgga aatcaaactt ataagcgtat ctcttaccgt
     2101 gtaccttggt cattatattt ttgtcataat agtatcggag ggctcgtgac aacttatcat
     2161 agttcatatt aggcttggac tttcgttctc cccaccgccg cgccacctca tcagggtcag
     2221 tcattttgaa ctctccgttg gtcccttccc acgttataca ggacgcatta gctgaatcgc
     2281 tgagcagttc aagaagaaat tgccacaatt gtatctgacc agagccgggg ttagcaagtc
     2341 ttgaagacgt aggtcccaag atctggtacg gatcgggttg gggtctctgt tctgtgttct
     2401 tgctaatggt ttgagcgccc cccaatgggg gtgacttatt caagccagag ttcatgttgt
     2461 tgccccacgc acctcttcta acagagtcat aagacgggtc accaaattta ttgaagcccc
     2521 ctctgtcgga gccccccatt cccccgcgtc caccccttcc accgcctcgg cctctcggct
     2581 catacccacc actggaccta ttgtatccac cccctccacc cccgccgcca ccaccactgc
     2641 cccccctgcc tctcccaccc ctgtcctgct gtccataccc accgctgcct ccgccgccag
     2701 attgatcttg atttccgtag cccccgccgg agcctccacc agaagacatt gaggactggt
     2761 cttgtccgta gtttccgccc ccaccgccgc cagagctgga attatattgg ttttgctgcc
     2821 cgtagccttg gggaggatta taggattgct gctgaccgta ggactgctgt tggcctccat
     2881 aactgggttg ttgggagtag ctcccactct gaggctggcc gtatgaacta ctttgggaac
     2941 tactcccgta actaccactt gtggaggatg gggctggttg ctgaccataa cctggatatg
     3001 aagattgttg accgtaggat gattgagaag attggcttga tccgtaaccg ccagtgcttc
     3061 cgtacccctg tggggttgac tgagtgccgt atcccgtatt ctggctttgc ccgtatgagc
     3121 tgtaagaact ttggccgtat ccggaggtgt cggtggactg actataccct gagtaggatt
     3181 gctgaccgta tggctggcta gactgttggc tgtatccctg tccgggttgt gtggggtatg
     3241 ccccataaga ttgcgttgct tgctgggtat aatcatttga tgccattccc cgggaagcgg
     3301 ccgcacttcc cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga
     3361 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     3421 tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     3481 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     3541 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     3601 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     3661 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     3721 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     3781 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     3841 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     3901 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     3961 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     4021 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta
     4081 ccgtcgactg cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta
     4141 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     4201 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     4261 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     4321 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     4381 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     4441 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     4501 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     4561 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     4621 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     4681 attactatta ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag
     4741 gtgggtctga aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac
     4801 agcaaaaact attcttaaac ctaccaagcc tcctactatc attatgaata attttatata
     4861 ccacagccaa tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa
     4921 taattcttgt tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa
     4981 gcttgtgtaa ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg
     5041 ttccagagat ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt
     5101 tccagagcaa ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag
     5161 gattcttgcc tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc
     5221 ctcaatagcc ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc
     5281 ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc
     5341 aaggaacaaa gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc
     5401 cttggtgggt gctactccta atggttcaat ttttactact ttatatttat ataattcact
     5461 tctccaattg tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta
     5521 atcctcatcc tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct
     5581 aatgatcttt gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac
     5641 tgccccttca cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc
     5701 cctgtaataa acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg
     5761 tatgtctgtt gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc
     5821 ttttctttta aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc
     5881 ctgatctctt acctgtccta taattttctt taattcttta ttcatagatt ctactactcc
     5941 ttgactttgg ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt
     6001 tttcttactt ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat
     6061 ctctatcctt tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc
     6121 ttctgatcct gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga
     6181 tgtttctaac aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat
     6241 atgttttaat ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga
     6301 tctaattctc ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc
     6361 gctagtcaaa atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc
     6421 ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc
     6481 cctgttcggg cgccactgct agagattttc cacactgact aaaagggtct gagggatctc
     6541 tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt
     6601 tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg
     6661 tctaaccaga gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc
     6721 tcgccactcc ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg
     6781 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     6841 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     6901 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     6961 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     7021 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     7081 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcgaccc
     7141 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     7201 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     7261 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7321 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7381 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7441 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7501 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7561 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7621 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7681 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7741 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7801 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7861 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7921 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7981 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     8041 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     8101 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     8161 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     8221 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     8281 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     8341 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8401 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     8461 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8521 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg g
//
