LOCUS       sequence-408065-        8520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, DDX3X_MLLT10 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8520)
  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 8520)
  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_DDX3X_MLLT10
FEATURES             Location/Qualifiers
     source          1..8520
                     /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(3281..3991,3992..3994,3995..3997))
                     /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(3323..3344)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3672..3691
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3931..3952
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3994..4003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4079..4282)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4108..4128)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4283..4586
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4843..4887)
                     /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    5072..5305
                     /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    5665..5782
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6274..6399
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6446..6703
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6705..7084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7115..7219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7220..7288,7289..8080)
                     /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(7438..7457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8251..8520,1..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 ccggttcatt
     1801 tgcgtacctg catgatctcc atcgaaggtg aaaaatgtgt cggcggcagc tgtgtaaagt
     1861 gtggatgacg gtggacaggc gtcaattgtt gagaatttaa taattgctga aattgctgct
     1921 gatgaattaa aatctgacgc tgctggtctg ataataatcc cagtccgcta gctgagttgg
     1981 taagcgtcgc tggcattggg tgtgttgcat ttggaggaac tgtggtatgc gttaacggtg
     2041 ttgggttctg gctcatggta acaggcgtct gcatgacacc attcagggcg cctacaatac
     2101 cgttgatagc cagctgatta ccgggtaaag caccaataat acctcccaca gctgagacag
     2161 cgggaatagt tgatgttacc ggttgcatac cgctagctaa cgcgcccacc gtcacaccat
     2221 ttacctgttg tactccgctg acacccgacc cttgctgtgc gggtgattgt cccgccgggg
     2281 gaggggtgga cagagcgctg ctactagatg tgctttgtcc cgaagaggtc agatcttggt
     2341 taagaacggg caatgaattg tctggcaaaa aggagtttcc aatatgtggt gacttcgacg
     2401 agtttaaaga atcagtcgtc ggtgcagtct gtgcactgaa agtatgaatt tgatgagacg
     2461 gcgacgggtt tgcggtgatt gtggggaaag gcacagaaag ctgagcatta agaagttgca
     2521 ggcgttcctt ctttgctgtc agatttttga tttgctcttc caggcgacgg ttctccactt
     2581 gtaactggtg cagagatttc aacataccca gaatgtcgga cggggttcct tgttccagca
     2641 gaaactgctg tccctcgctc cattgacgtt ccagcagttg ctcaatcgat gctgccacgg
     2701 gcggcaaatt ttctaatgaa ctgtttccag gttgatcgta acggatttgt aacgaggata
     2761 ccggagagcg aggtgacaac gaaccacggg gggaggaacc acgtcctact aaattacgtg
     2821 aattatcgcc cagatcctgg tctgtctggt tgttctcact ttgtgcgatt aaggcagcca
     2881 tagaactatt ggagcggttc ccatacatgt gggaaggagc ctgagaaagt gaagatcccg
     2941 aaagggtgtt ggcttgcgtg gttgcaaccg cgggggccgc agaggagaca gcagagggag
     3001 acaaagcgcc aacctgttga aggtgaccgg agctctgttg gggaaggtgg ctagaactaa
     3061 cgggagtcga actgcctgag ccagacacca cattagggaa gctaacagcc acatcgttcg
     3121 aattgtaaat actagaagct gtcgaacccc cagattggtt gtcggaggag ttcagatcta
     3181 agccggcgaa ctgttggtcc agacccaggg cattctcaac agccacgtga ctcattcccc
     3241 gggaagcggc cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc
     3301 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     3361 tggggtcttt gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     3421 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     3481 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     3541 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     3601 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     3661 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     3721 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     3781 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     3841 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     3901 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     3961 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg
     4021 cccgcggtac cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag
     4081 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     4141 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     4201 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     4261 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     4321 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4381 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4441 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4501 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4561 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4621 ccgtaattga ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg
     4681 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
     4741 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
     4801 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
     4861 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
     4921 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
     4981 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
     5041 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
     5101 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
     5161 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
     5221 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
     5281 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
     5341 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
     5401 taattcactt ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc
     5461 catgttctaa tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc
     5521 ataatcccta atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg
     5581 tattactact gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt
     5641 tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc
     5701 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc
     5761 aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag
     5821 atgttcagcc tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc
     5881 tactactcct tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg
     5941 ctgtgctttt ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt
     6001 gtcttttatc tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga
     6061 tctaagttct tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca
     6121 gccttctgat gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc
     6181 ccatactata tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc
     6241 ccatcgcgat ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt
     6301 ctagcctccg ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc
     6361 cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc
     6421 tttcaagtcc ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg
     6481 agggatctct agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa
     6541 ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct
     6601 cagatctggt ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga
     6661 tctgagggct cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg
     6721 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     6781 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     6841 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     6901 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     6961 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     7021 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     7081 tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7201 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7261 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7321 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7381 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7441 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7501 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7561 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7621 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7681 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7741 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7801 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7861 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7921 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7981 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8041 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8101 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8161 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     8281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     8401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
//
