LOCUS       sequence-407534-        8655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MBNL1_MEIS2 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8655)
  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 8655)
  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_MBNL1_MEIS2
FEATURES             Location/Qualifiers
     source          1..8655
                     /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(3416..4126,4127..4129,4130..4132))
                     /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(3458..3479)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3807..3826
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4066..4087
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4129..4138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4214..4417)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4243..4263)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4418..4721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4978..5022)
                     /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    5207..5440
                     /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    5800..5917
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6409..6534
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6581..6838
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6840..7219
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7250..7354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7355..7423,7424..8215)
                     /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(7573..7592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8386..8655,1..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 ccggttcact
     1801 gtgcgtgaat atccataact tgaccgccca catttggatc aacgctattc agcattgttg
     1861 gactttgagc ggacatggtc attccgggat gagttggagg tcctccatgc atcatcattg
     1921 cgggatgatg tggatggctc ggcaagtatg aatgcatcgg cggcccatgg cgtaattgcg
     1981 tggggtgtgg cgtcatttgg ggaggggtgt acgacggctg agccattgac atccccatgg
     2041 gaccgccctg actaacataa tcgccgggca tactctgaag accggcagga cggattccca
     2101 tgtgttgttg cccgtccaaa acaaacgacc ccattggttg cccctcaggg gagtaggcgg
     2161 cgccttggga tacggcgcga ttactctggt caatcattgg ctgtacgatg cggcggcgag
     2221 cattaataaa ccagttgttg acttgaagga tggtcaggcc ggtatcttgt gctaattgtt
     2281 ttttctgctc ttcagacggg tacggatgcg taaggtgttg gaacaaccaa gcgcgcataa
     2341 tgttggttgc tactttcgga aaaatgccgc gttttttttg acgtttttta tccttatctg
     2401 ggtcgtcatc atcaccggtg cccggtgatg ccacggaatt gtcaagccca tctccctgtt
     2461 cagatgagtt gtctcccgac tgggatgcat gaccccccga agaaggaccg ggtgttcccg
     2521 cgctgtgtgt cgaggtagcg tcatcgtgat cgcgccagga actaggatta tggtcggcaa
     2581 ggttcgtgct cgaccccgaa agttcctcgt gatcagattt gctactgcca tcgcgttcat
     2641 caataaccag gtcaatcggc attttgcctt taaggcaaga gatataacgg tggcagaaat
     2701 tatcacataa ctcatggact ttctccaatt ccagcaggtg aaaacgcaat acttgaattg
     2761 cttggatcat taagttatct aattcggggt ttgatgagaa cagcggcttc tccgcacgga
     2821 cttgcttcgc aaagacggca atatcttcgt tgaacgaatc tgaactgcag acatcgcccc
     2881 ccgcaacccc tggctcacgt ggtgtgcaag tagcaagttc acacttttca aagaccaagg
     2941 caagtaaggg gaaaagggga tggccgtaaa tggcgtcttt gtcgcgcttc agcgcgtcgt
     3001 ttaccgcact ccccatggac gctggcataa cattgggatg cggtgcatgc gccccgtaat
     3061 gctgcgtagc gtgtaagggg ggaccatggt tcaagtgatg tactggtggg atgggacggg
     3121 gcgcgtgagg gtcgccgtac atggacgctg gaactcctac accatccatc ccaccgtagt
     3181 ggggtaattc gtcatagact ggttgcaggg gcgcacctgg catcatggca ttagccagct
     3241 gcatttgttg tgctaacatg gccatgttct tttgttgaat caaattatta cggccattaa
     3301 tttctaattg tgttttaaga tgaggaggtg gatgtaaata tttgcaattt tcgcggctac
     3361 aacgtcccat tccccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt
     3421 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     3481 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     3541 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     3601 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     3661 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     3721 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     3781 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     3841 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     3901 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     3961 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     4021 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     4081 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     4141 ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc
     4201 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     4261 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     4321 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     4381 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     4441 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4501 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4561 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4621 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4681 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4741 tatgaactaa tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga
     4801 gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
     4861 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
     4921 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
     4981 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
     5041 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
     5101 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
     5161 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
     5221 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
     5281 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
     5341 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
     5401 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
     5461 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
     5521 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt
     5581 tttactaaac ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc
     5641 tgccatctgt tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat
     5701 gtcactatta tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc
     5761 tttccaaagt ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     5821 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     5881 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     5941 tactgctgtc ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc
     6001 tttattcata gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga
     6061 agatcctgct gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc
     6121 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     6181 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     6241 cagtatttgt ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc
     6301 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     6361 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     6421 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     6481 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     6541 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     6601 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     6661 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     6721 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     6781 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     6841 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6901 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6961 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     7021 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     7081 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     7141 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     7201 ctagtcaata atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7321 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7501 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7681 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7741 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7981 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     8041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     8101 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     8161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     8221 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8281 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8341 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8401 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8461 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8521 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8581 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8641 gtcgtgtctt accgg
//
