LOCUS       sequence-406642-        7203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, DDX5_ZNF521 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7203)
  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 7203)
  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_DDX5_ZNF521
FEATURES             Location/Qualifiers
     source          1..7203
                     /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(1964..2674,2675..2677,2678..2680))
                     /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(2006..2027)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2355..2374
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2614..2635
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2677..2686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2762..2965)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2791..2811)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2966..3269
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3526..3570)
                     /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    3755..3988
                     /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    4348..4465
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4957..5082
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5129..5386
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5388..5767
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5798..5902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5903..5971,5972..6763)
                     /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(6121..6140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6934..7203,1..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 ccggttcagc
     1801 attgttttgt tttcttcttg ttactcttga acccgcgcgc ggctccttgg ccaaagcaaa
     1861 tagttgaagg gatgccacga tcacggccgc gatcacggtc cgaactgtaa ccggacattc
     1921 cccgggaagc ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca
     1981 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     2041 cgttggggtc tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca
     2101 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     2161 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     2221 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     2281 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     2341 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     2401 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     2461 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
     2521 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     2581 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     2641 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc
     2701 gggcccgcgg taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac
     2761 gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     2821 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     2881 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     2941 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     3001 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     3061 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     3121 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     3181 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     3241 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     3301 accccgtaat tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct
     3361 cggggttggg aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac
     3421 tatagaaagt acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa
     3481 taattttata taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt
     3541 atctaattcc aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa
     3601 ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg
     3661 attccaaatc tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt
     3721 gcaaatgagt tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct
     3781 ttccacagcc aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag
     3841 atgctgttgc gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa
     3901 taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc
     3961 tcccaagaac ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac
     4021 tcttctcttt gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt
     4081 atataattca cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt
     4141 ttccatgttc taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt
     4201 tccataatcc ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc
     4261 ttgtattact actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg
     4321 atttctgctg tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa
     4381 ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc
     4441 cccaatcccc ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt
     4501 aagatgttca gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga
     4561 ttctactact ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc
     4621 ttgctgtgct tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt
     4681 ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa
     4741 tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct
     4801 acagccttct gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct
     4861 tgcccatact atatgtttta atttatattt tttctttccc cctggcctta accgaatttt
     4921 ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc
     4981 cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct
     5041 tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt
     5101 cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt
     5161 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     5221 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     5281 gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca
     5341 ggatctgagg gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg
     5401 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     5461 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     5521 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     5581 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     5641 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     5701 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     5761 caatgtcgac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5821 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5881 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6001 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6061 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6181 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6301 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6481 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6721 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6781 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6841 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7081 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7201 cgg
//
