LOCUS       sequence-407536-        8505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MEF2D_DAZAP1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8505)
  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 8505)
  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_MEF2D_DAZAP1
FEATURES             Location/Qualifiers
     source          1..8505
                     /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(3266..3976,3977..3979,3980..3982))
                     /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(3308..3329)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3657..3676
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3916..3937
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3979..3988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4064..4267)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4093..4113)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4268..4571
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4828..4872)
                     /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    5057..5290
                     /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    5650..5767
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6259..6384
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6431..6688
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6690..7069
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7100..7204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7205..7273,7274..8065)
                     /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(7423..7442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8236..8505,1..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 ccggttcagc
     1801 gacgataggg atggaagccc tgaacattgt ggttctgccc acggccaaaa ccactaccgc
     1861 ctgcaggcgg accccctgag ccaggaacag aggggccacc ataactcgga ggttgctggg
     1921 atggatcact gaagccttgg ccgaaacctg ataagtcctg tccgtaaccg gcatactgcc
     1981 cataagggaa gtctggctgc gctgttggtg gcttggacat gtcgggggca gcctgactag
     2041 gaggcggcgg aaaggccaaa ggtgccgcac ctggggtagc tggtggaggt gggacacctg
     2101 gcggggcaaa ttggtcgggt ggtggcgggg gtggcccata gccaaaacta aactgcgggg
     2161 gagcgccata cccctgaggg aacccctggg gaggtggaaa ccctcctggg ggcgtagaga
     2221 cgatataaga cgtaaatggt ggtggaggcg gcggcgcgcc acgtcctgcg ggcgggggac
     2281 catagccccc gatggcttgt ccagcgggta cccacattcc ctgtggaccg tagccttgct
     2341 gccaggtcgg aggtggttga cccgcccaac catttgcggc gttgggaacg acacgtgacc
     2401 cccactgact tgcaccgggt tgaccggggg cctgactctt tgagtcgcgt ggttcggcac
     2461 gcttaacttc aacctttttc cccatgatgt cgtggaagtg catgttcacg gcttggtcca
     2521 ccgattgctc gtcctcaaaa gtaatgaatc cgaagccgac gggcgacggg caagcaccgt
     2581 tagcggagtt caaatcccct cctaacatcg ctcctgcact agctggacgt tgaggtaacc
     2641 caggggacac gctgttgcgt tgtaatgcag gctgctgcgg tgacagcaaa cgaggatcgg
     2701 taaggcttga agtgaccagc gacggggtca ccaaactccc tgatggatta ctaaattgaa
     2761 gggaactctg gttactaact ggtacggtta cgggcatcgc aaagttggga gctggaacgg
     2821 tagatccgta gcgacggaat aacccatcca gctcctcact tgcgcggcgg tatttatctt
     2881 ccagcagtgg gctctgttcc aatgaatcct cgccatcggg ctcaggtgag tcgcaaccat
     2941 taaacccctt cttacgcagc gtctcgatga tgtcagcatt cgtacggctc tcgtgaggct
     3001 cattgtactc ggtgtatttc aacagcactt tgtccatgtc ggtcgatgca tattggaaca
     3061 acttgtttga gtgattaaag atgatcaagg cgatttcaca atcgcacaaa acggataatt
     3121 cgtaggcttt tttcatcaac ccaaacttgc gcttagtgaa cgtcacttga cggttgcgtt
     3181 cgtccgtaat gcgctgaatc tggatctttt tacgtcccat tccccgggaa gcggccgcac
     3241 ttcccccgga gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg
     3301 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     3361 gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     3421 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     3481 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     3541 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     3601 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     3661 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     3721 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     3781 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     3841 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     3901 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     3961 gctcctcgcc cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg
     4021 actgcagaat tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga
     4081 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     4141 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     4201 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     4261 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4321 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4381 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4441 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4501 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     4561 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     4621 attaataact agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt
     4681 ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa
     4741 aactattctt aaacctacca agcctcctac tatcattatg aataatttta tataccacag
     4801 ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc
     4861 ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt
     4921 gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag
     4981 agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga
     5041 gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct
     5101 tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat
     5161 agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac
     5221 cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa
     5281 caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt
     5341 gggtgctact cctaatggtt caatttttac tactttatat ttatataatt cacttctcca
     5401 attgtccctc atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc
     5461 atcctgtcta cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat
     5521 ctttgctttt cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc
     5581 ttcacctttc cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta
     5641 ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc
     5701 tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct
     5761 tttaaaattg tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc
     5821 tcttacctgt cctataattt tctttaattc tttattcata gattctacta ctccttgact
     5881 ttggggattg tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt
     5941 acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat
     6001 cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga
     6061 tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc
     6121 taacaggcca gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt
     6181 taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat
     6241 tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt
     6301 caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc
     6361 aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt
     6421 cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
     6481 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
     6541 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
     6601 cagagagacc cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca
     6661 ctccccagtc ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg
     6721 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     6781 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     6841 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     6901 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     6961 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     7021 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg
     7081 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7141 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7201 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7261 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7321 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7381 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7441 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7501 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7561 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7621 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7681 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7741 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7801 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7861 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7921 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7981 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     8041 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     8101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     8221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     8281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     8341 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     8401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgg
//
