LOCUS       sequence-408121-        8820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MEF2D_BCL9 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8820)
  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 8820)
  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_BCL9
FEATURES             Location/Qualifiers
     source          1..8820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..38
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    106..240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(130..149)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     184..203
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(246..441)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      260..395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    500..521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(536..541,542..559,560..566)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    574..590
                     /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     579..601
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     598..614
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     598..614
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     957..976
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    975..1030
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1076..1095
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3129..3839,3840..3842,3843..3845))
                     /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(3171..3192)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3520..3539
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3779..3800
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3842..3851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3927..4130)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3956..3976)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4131..4434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4691..4735)
                     /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    4920..5153
                     /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    5513..5630
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6122..6247
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6294..6551
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6553..6932
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6963..7067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7068..7136,7137..7928)
                     /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(7286..7305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8099..8687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8588..8607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
       61 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc agacatgata
      121 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
      181 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
      241 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
      301 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
      361 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
      421 gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac tggaaagcgg
      481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      601 gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga tctccataag
      661 agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct ggctagtaaa
      721 acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa
      781 aaaatctaac ctcaagtcaa ggcttttcta tggaataagg aatggacagc agggggctgt
      841 ttcatatact gatgacctct ttatagccaa cctttgttca tggcagccag catatgggca
      901 tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt tgccctccca
      961 tatgtccttc cgagtgagag acacaaaaaa ttccaacaca ctattgcaat gaaaataaat
     1021 ttcctttatt agccagaagt cagatgctca aggggcttca tgatgtcccc ataatttttg
     1081 gcagagggaa aaagatctgc tagctcgagc agctgaagct ttattgaggc ttaagcagtg
     1141 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
     1201 tacaggcaaa aagcagatct tgtcttcttt gggagtgaat tagcccttcc agtcccccct
     1261 tttcttttaa aaagtggcta agatctacag ctgccttgta agtcattggt cttaaaggta
     1321 ccgtaatctt atctagatca aaacatcatg tttccgggat tgcccggtcc ttgtgaaaac
     1381 ccacccattc ccacatctgc tgccatgcca cggggacgca tctgaggagg gatcataatg
     1441 ttttgttggg gtcccatcat tccctgcatg ctcatcatca tacctggact acccacaggt
     1501 ccgggatgcg tatacagtcc agcaggtcca cgatctttgc cgggaatcat ccccacggcg
     1561 gcggctggat tactcataag cgtgggttgc ccaggcatgg tcgattgcgc cggagacatc
     1621 atacgatgat gtggtcccat catcccctgc tgcaagaacg ctggcggacg catagggttg
     1681 tggcctggca tacttggagc cgtaccaagt gggatgtcgg gcgtcccgac tgggcccgga
     1741 ccacccattc ccggtaaagc caaacccata cggggtgcct gctctcccat cattccttgc
     1801 atgtgggaaa accctggtcc gggcccctgg ggttgtttgc gaccagggac ctcgccgcga
     1861 gggaagtact gcagcgtctg tgagggtttc tctgacggaa taatacgaga aagatcaaac
     1921 tcaggaatgc cggtagcacc gggacgaatt acttcttgca aatcgggatc agtaaacacg
     1981 gatggcatgc tgttccccaa gacggtgaaa ctatccggcc ctccgggacc tcccggttta
     2041 cacagcgcag catcagctga actctgcggc aagtttgatg ggcgacccag tggcccttca
     2101 cccggaaatc ccatgccacc aggaaatgag ccttggcccc cggagggccc gttgtgggga
     2161 aaggggacct gttgcggtgg tgactgcacc ggaggaaagc cctgtggaaa gcccatgcgc
     2221 ccctgaggga ccataggagg ttcctgtgat ccatgtccca taattggatt atgactcatt
     2281 aagcctggcc ccattggcac tccgtgggga gggatattcg ggccaacggc gtttggcgag
     2341 ggcatttgat tggaatggct taatggctgc gtcattccca taggggaaag tgtgggcata
     2401 ggaactactg ggttaggacc cactggggat gggcaagcgc catttgcgct attcaagtct
     2461 ccaccaagca ttgcccccgc cgaggctggg cgttgcggta aacctggtga gacactatta
     2521 cgttgaagtg cgggctgctg tggcgacaac aagcgtgggt ccgtaaggga cgaggttacc
     2581 agtgagggag tgactaagct gccagacggg ttactgaact gaaggcttga ttggttagag
     2641 accgggacgg tcacgggcat cgcgaagttg ggtgccggaa ctgtagatcc atagcgacgg
     2701 aacagtccgt caagctcttc ggacgcacgg cggtacttat cttcaagcag tggggattgt
     2761 tcaagcgaat cttcgccatc gggttcgggg ctatcacacc cgttgaaccc cttcttgcgc
     2821 aacgtctcga taatgtctgc attggtgcga ctctcgtggg gctcattgta ctcagtgtac
     2881 ttcaacagca ccttatccat atctgttgaa gcgtactgga aaagtttatt gctgtgattg
     2941 aaaatgatta aagcgatctc gcaatcgcat aatactgaca gctcgtaggc cttcttcatc
     3001 aaaccgaact tgcgcttcgt gaacgtcacc tggcgattgc gctcatccgt aatacgttgg
     3061 atctggatct tcttgcggcc cattccccgg gaagcggccg cacttccccc ggagcctccg
     3121 cttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3181 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctgggtgctc
     3241 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3301 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     3361 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     3421 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     3481 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     3541 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     3601 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     3661 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     3721 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     3781 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     3841 accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcggttc
     3901 actaaacgag cccggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc
     3961 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     4021 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     4081 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     4141 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     4201 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     4261 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     4321 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     4381 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     4441 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagcacgc
     4501 gttcgaagct tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg
     4561 aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta
     4621 ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa
     4681 accaattcca caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt
     4741 gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc
     4801 tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac
     4861 tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca
     4921 ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga
     4981 tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt
     5041 tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg
     5101 ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca
     5161 ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg
     5221 gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc
     5281 ctccttatgg tgttttacta aacttttcca tgttctaatc ctcatcctgt ctacttgcca
     5341 cacaatcatc acctgccatc tgttttccat aatccctaat gatctttgct tttcttcttg
     5401 gcactacttt tatgtcacta ttatcttgta ttactactgc cccttcacct ttccagagga
     5461 gctttgctgg tcctttccaa agtggatttc tgctgtccct gtaataaacc cgaaaatttt
     5521 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
     5581 ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga
     5641 atactgccat ttgtactgct gtcttaagat gttcagcctg atctcttacc tgtcctataa
     5701 ttttctttaa ttctttattc atagattcta ctactccttg actttgggga ttgtagggaa
     5761 tttccaggtc tgaagatcct gctgcttgct gtgctttttt cttacttttg ttttgctctt
     5821 cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat
     5881 agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg
     5941 gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccagtattaa
     6001 ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct
     6061 ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac
     6121 tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac
     6181 tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc
     6241 gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga
     6301 gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag
     6361 acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt
     6421 ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca
     6481 ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca gtcccgccca
     6541 ggccacgcct cccatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     6601 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     6661 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     6721 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     6781 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     6841 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     6901 ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt cggggaaatg
     6961 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7021 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7081 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7141 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7201 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7261 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7321 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7381 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7441 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7501 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7561 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7621 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7681 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7741 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7801 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7861 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7921 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7981 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     8041 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     8101 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8161 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8221 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8281 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8341 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8401 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8461 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8521 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8581 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8641 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8701 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8761 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
//
