LOCUS       sequence-406577-        8013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, ASCC1_MICU1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8013)
  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 8013)
  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_ASCC1_MICU1
FEATURES             Location/Qualifiers
     source          1..8013
                     /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(2774..3484,3485..3487,3488..3490))
                     /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(2816..2837)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3165..3184
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3424..3445
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3487..3496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3572..3775)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3601..3621)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3776..4079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4336..4380)
                     /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    4565..4798
                     /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    5158..5275
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5767..5892
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5939..6196
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6198..6577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6608..6712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6713..6781,6782..7573)
                     /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(6931..6950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7744..8013,1..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 gcttcgggag ggcaaagtcc caggcagtct cttgtgcgca cttccacata gcttgcataa
     1861 ggcgagtaaa ccccatgtcc ttaggttttt ccagccctcg catcagtcgc tgcttcataa
     1921 ttgaaacaaa ctccttgttg gaaagctcgc catttccgtc acagtcgaaa agcgcgaaga
     1981 ctacgtcaca aacgtgatca ctcaactcca ctttggcgac agtccgggct acttgctgca
     2041 tagtcacttt atcgagagat gcacctgcca tgtggtagaa ggagagagcc gtatctacat
     2101 catttatatt cttcaggaac gtgaaaaagt tctctacctc ttgaaaggtc aaacccatgc
     2161 tacatttagc caggacttcc tcctgaaatc tcaggaaccc ctcttggacc tcgacttcat
     2221 tcaagaagaa ggccaggaag tgtgtgaagg gctgctttct cctgaaggtg tccagcaaaa
     2281 catcaattct cgtccttgcg gagatcacac cgtttctatg ctgcccagtt ataacgattt
     2341 ccccatcctg accgggtttg ggtatagata tggatgtttt tgtctccatt tcgatttttt
     2401 tcctagtgtc ccctcgtttc cccacgatat gtcttgattt ccaggtataa atattctggc
     2461 agtccaaagt aattttctga aatccacaat cgtttgaggt attaagatgt ataagattgt
     2521 acaacagaga gggggctcgc aaggttgagc gaaaaccttg cggtgtctgc tccacttcat
     2581 aggcgtcgca cggttcgtca gcgcactcca ttgacccttg gtagaaatct tcctcatcct
     2641 cttcgtgctg gtaagtttgc tcttgcacgg gattcttccg gtagttgcga ccgtcgattc
     2701 gaatgagttg ggggcgcaag acttccattc cccgggaagc ggccgcactt cccccggagc
     2761 ctccgcttcc gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     2821 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg
     2881 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     2941 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     3001 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     3061 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3121 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3181 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     3241 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     3301 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3361 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     3421 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     3481 tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc
     3541 ggttcactaa acgagcccgg ttcactaaac gagctctgct tatatagacc tcccaccgta
     3601 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     3661 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     3721 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     3781 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     3841 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     3901 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     3961 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4021 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4081 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4141 cacgcgttcg aagcttgagc tcggtcccct cggggttggg aggtgggtct gaaacgataa
     4201 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     4261 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     4321 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     4381 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     4441 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     4501 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     4561 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     4621 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     4681 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     4741 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     4801 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     4861 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     4921 atctcctcct tatggtgttt tactaaactt ttccatgttc taatcctcat cctgtctact
     4981 tgccacacaa tcatcacctg ccatctgttt tccataatcc ctaatgatct ttgcttttct
     5041 tcttggcact acttttatgt cactattatc ttgtattact actgcccctt cacctttcca
     5101 gaggagcttt gctggtcctt tccaaagtgg atttctgctg tccctgtaat aaacccgaaa
     5161 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     5221 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg
     5281 gatgaatact gccatttgta ctgctgtctt aagatgttca gcctgatctc ttacctgtcc
     5341 tataattttc tttaattctt tattcataga ttctactact ccttgacttt ggggattgta
     5401 gggaatttcc aggtctgaag atcctgctgc ttgctgtgct tttttcttac ttttgttttg
     5461 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
     5521 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
     5581 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagt
     5641 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
     5701 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
     5761 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
     5821 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
     5881 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
     5941 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
     6001 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
     6061 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
     6121 gtacaggcaa aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc
     6181 gcccaggcca cgcctcccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     6241 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     6301 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     6361 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     6421 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     6481 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     6541 attgattact attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg
     6601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7861 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7981 tgctgccagt ggcgataagt cgtgtcttac cgg
//
