LOCUS       sequence-407507-        8172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, ETV6_C7ORF72 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8172)
  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 8172)
  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_ETV6_C7ORF72
FEATURES             Location/Qualifiers
     source          1..8172
                     /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(2933..3643,3644..3646,3647..3649))
                     /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(2975..2996)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3324..3343
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3583..3604
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3646..3655
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3731..3934)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3760..3780)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3935..4238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4495..4539)
                     /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    4724..4957
                     /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    5317..5434
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5926..6051
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6098..6355
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6357..6736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6767..6871
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6872..6940,6941..7732)
                     /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(7090..7109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7903..8172,1..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 aatcaatggt ttctttgtct ttcttgttaa agggattata aggacgcacg gtagtgacta
     1861 aacgcgacaa tggggcttga tgacgaaaca ggtctacggc catctccttt tgaaatacac
     1921 gatgtaactc ggaatccggc gacggtgtgg tagatgggat gttagagttc gctggatgtg
     1981 tggcagtgaa atgcactttc cccgtgtaac ccggaatgtt cgctgcacgg ctagtgttag
     2041 tatataacgg tttcgagtag cgctgtttgg ctaaggacgc aatgtcgccc agcggattgt
     2101 ctacgtcgtc cgcgtttgta ctttgaacgt ggcccgtgta taacggaatg tacttgctac
     2161 ggctactcga tgatacaaaa ttgatgggtt taacaggcgc aagaaaacta cgtgtctgaa
     2221 aacgatccca taactcaccc accatctgag taatggctgg tactccctta tagcgtttta
     2281 acgtaaaaca ctggctgatg gggtcggcgg tcttctctaa agtagtctct tctgggacca
     2341 gcttcggcgg aagcatttta tcccacggaa cgtcttcgta tgctttctgt gtcgaggatt
     2401 tataaaagct atctttggca ttacactttt tgttggtctg taccagaggc tctccttcgt
     2461 cggtgcatgt ggcttcttca tcaaaagtaa agatgtgaga aagggaacgt ccctcatgat
     2521 gaggttgcag tggagtcact ttaaggggac tagtccaccc actatgaggg gaacgataac
     2581 ggaaatcttc ctttgtcagc agcaaaagag ccttaccatt catttcaaaa gtattggaat
     2641 cgatgggacg aagcgagaac tcattttctg cccatttcag ccattgagct acgtcatcac
     2701 gactccagta gattggttgc aaacgcaagt gcgctggcag gcggattgaa tcttcctcca
     2761 tacgaagtgc gcgtggaacc ggcacgtgaa ggggtgttga gctagcgtac gatggaaccg
     2821 gagattcggg cggggtatag gaaatgcgct cttgcttgat actacactga gcgggcgtct
     2881 cgctcattcc ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca
     2941 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     3001 cgcgcttctc gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg
     3061 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     3121 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     3181 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     3241 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     3301 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     3361 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     3421 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     3481 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     3541 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     3601 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg
     3661 gtggatcccg ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt
     3721 tcactaaacg agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     3781 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     3841 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     3901 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     3961 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4021 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4081 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     4141 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     4201 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     4261 gaactaatga ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct
     4321 cggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac
     4381 tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat
     4441 cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact
     4501 tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc
     4561 ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca
     4621 ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac
     4681 agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt
     4741 taggtatctt tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag
     4801 ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat
     4861 accagacaat aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct
     4921 tcctgctgct cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct
     4981 ctgcaccact cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac
     5041 tttatattta tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt
     5101 actaaacttt tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc
     5161 catctgtttt ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc
     5221 actattatct tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt
     5281 ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg
     5341 tttttgtaat tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc
     5401 actgtacccc ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac
     5461 tgctgtctta agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt
     5521 attcatagat tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga
     5581 tcctgctgct tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa
     5641 agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt
     5701 attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag
     5761 tatttgtcta cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag
     5821 ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa
     5881 ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc
     5941 atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc
     6001 ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg
     6061 tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac
     6121 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
     6181 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
     6241 agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc
     6301 ttatatgcag gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg
     6361 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     6421 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     6481 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     6541 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     6601 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     6661 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     6721 gtcaataatc aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6781 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6841 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6901 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6961 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     7021 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     7081 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     7141 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     7201 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     7261 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7321 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7381 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7441 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7501 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7561 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7621 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7681 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7741 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7801 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     8041 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8161 gtgtcttacc gg
//
