LOCUS       sequence-407476-        8310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, TRIOBP_RBFOX2 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8310)
  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 8310)
  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_TRIOBP_RBFOX2
FEATURES             Location/Qualifiers
     source          1..8310
                     /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(3071..3781,3782..3784,3785..3787))
                     /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(3113..3134)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3462..3481
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3721..3742
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3784..3793
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3869..4072)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3898..3918)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4073..4376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4633..4677)
                     /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    4862..5095
                     /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    5455..5572
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6064..6189
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6236..6493
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6495..6874
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6905..7009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7010..7078,7079..7870)
                     /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(7228..7247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8041..8310,1..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 ccggttcaat
     1801 aaggtgcgaa gcggctgtaa ccaccgcggt acaacgaggc tacagccccc acaccatagc
     1861 ttgcggcagg cgcaagagcg tgatatgggt cagcggtgta gacgcgtccg tagccatcac
     1921 tgtatgctgc cgctgctgct gccgcagcgg tggccgcagt tgctgtagcc ggctgcgcat
     1981 aacggtaagc agcgtagccc ccataaaggt cagctccata gaacccatcc tggtagacca
     2041 caccaggata tgctgggatg gctgttggag gaacggcacg tactgcacca tatactgtgc
     2101 gtccgcgtcc gcgcaaatgt gcgccacgaa acgctgcagc agtcgtcgcg gcggttgggt
     2161 atgggaagcc tggcactaat ggcagggaaa taagcggaat ataagtatta atccctccgc
     2221 ggccggataa tggtacggca gcatcattac caagggatac atctgcctgg aagctgctcg
     2281 ctgcgtaaag ctccgggcca tagacagcgc caacaaccgg agacaatttc catccattcg
     2341 catacggtgt taccattttt ttgtttgtca tgacacgcgc agtcgcattg ttcacttcga
     2401 ttttgcgacc ctcaaccacg gtgccgtgca acttttcgcg cgcacggtcg gcatccgctg
     2461 agttttcgaa cgttacgaaa ccaaaccctt tagaaccgcg ctcgttaaag ataatctcaa
     2521 catcaagaat ctttccaaat tgtccgaaca tttgacgcaa atcagggtcg cgaaaacgaa
     2581 aagggatgtt actaacatga agacgcttag gagtcgattt agactcgctg ttttcactac
     2641 tctgtgtttg ggattgctgc ccgtcagttt gggctccacc ctcggtcgta agcgacccgt
     2701 tctgtgttga cggggaattg ctactctgtt caccgtgagc ctgggttgac ccgtacaagg
     2761 taagattgtg ttctccagtt tggcccgcat aatcctgagt gtggggcaca ccgtactccg
     2821 tcgggatgcc gttttggggc ggagggggaa agggaatagt cgtgaagggt tggaccatag
     2881 catcaggggt cgtagtcggc tcctgattgc cttggtagcg ggctccgtga gcctcctccg
     2941 gatgaaagca gttttgacaa cggttttgag tcagaatgtt cgcctcgaaa tgctcacata
     3001 acgcgtctcc aggtacttcc tccattcccc gggaagcggc cgcacttccc ccggagcctc
     3061 cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     3121 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc
     3181 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     3241 agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct
     3301 tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact
     3361 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     3421 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     3481 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     3541 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
     3601 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
     3661 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     3721 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     3781 tcaccatggt ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt
     3841 tcactaaacg agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac
     3901 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     3961 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     4021 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     4081 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4141 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4201 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4261 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4321 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4381 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac
     4441 gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     4501 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     4561 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     4621 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     4681 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     4741 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     4801 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     4861 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     4921 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     4981 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     5041 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     5101 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     5161 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     5221 tcctccttat ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc
     5281 cacacaatca tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct
     5341 tggcactact tttatgtcac tattatcttg tattactact gccccttcac ctttccagag
     5401 gagctttgct ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt
     5461 ttgaattttt gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc
     5521 tactattctt tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat
     5581 gaatactgcc atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat
     5641 aattttcttt aattctttat tcatagattc tactactcct tgactttggg gattgtaggg
     5701 aatttccagg tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc
     5761 ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca
     5821 atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga
     5881 tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt
     5941 aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt
     6001 ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat
     6061 actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt
     6121 actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg
     6181 tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta
     6241 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     6301 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     6361 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     6421 caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc
     6481 caggccacgc ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     6541 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     6601 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     6661 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     6721 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     6781 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     6841 gattactatt aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa
     6901 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7921 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7981 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8161 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8281 tgccagtggc gataagtcgt gtcttaccgg
//
