LOCUS       sequence-407530-        8157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MEX3D_DCTN6 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8157)
  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 8157)
  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_MEX3D_DCTN6
FEATURES             Location/Qualifiers
     source          1..8157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(115..219)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             631..888
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    935..1060
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1552..1669
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2029..2262
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2447..2491
                     /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"
     enhancer        2748..3051
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3052..3255
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3206..3226
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3331..3340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3337..3339,3340..3342,3343..4053)
                     /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(3382..3403)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3643..3662)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3990..4011
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5424..5443)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5489..5544
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5543..5562)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5905..5921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5905..5921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5918..5940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5929..5945
                     /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)."
     promoter        complement(join(5953..5959,5960..5977,5978..5983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5998..6019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6078..6273
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6124..6259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6279..6413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6316..6335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6370..6389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6481..6498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6652..7240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6732..6751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7411..8157,1..45,46..114))
                     /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     8034..8053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
       61 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      241 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg gaggcgtggc ctgggcggga ctggggagtg
      661 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
      721 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      781 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      841 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      901 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      961 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     1021 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     1081 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     1141 acatatagta tgggcaagca gggagctaga acgattcgca gttaatactg gcctgttaga
     1201 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     1261 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     1321 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     1381 gaaaaaagca cagcaagcag caggatcttc agacctggaa attccctaca atccccaaag
     1441 tcaaggagta gtagaatcta tgaataaaga attaaagaaa attataggac aggtaagaga
     1501 tcaggctgaa catcttaaga cagcagtaca aatggcagta ttcatccaca attttaaaag
     1561 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     1621 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     1681 cagggacagc agaaatccac tttggaaagg accagcaaag ctcctctgga aaggtgaagg
     1741 ggcagtagta atacaagata atagtgacat aaaagtagtg ccaagaagaa aagcaaagat
     1801 cattagggat tatggaaaac agatggcagg tgatgattgt gtggcaagta gacaggatga
     1861 ggattagaac atggaaaagt ttagtaaaac accataagga ggagatatga gggacaattg
     1921 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     1981 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     2041 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     2101 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     2161 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     2221 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     2281 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     2341 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     2401 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     2461 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     2521 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     2581 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     2641 ccacctccca accccgaggg gaccgagctc aagcttcgaa cgcgtgctag ttattaatag
     2701 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2761 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2821 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2881 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2941 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3001 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3061 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3121 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3181 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3241 tatataagca gagctcgttt agtgaaccgg gctcgtttag tgaaccgaat tctgcagtcg
     3301 acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
     3361 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3421 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3481 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     3541 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3601 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3661 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3721 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3781 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3841 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3901 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     3961 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4021 gggatcactc tcggcatgga cgagctgtac aagggcggaa gcggaggctc cgggggaagt
     4081 gcggccgctt cccggatgcc ctctagcctt gggcaaccag atgggggagg tggaggtggt
     4141 ggtggcggcg gcggggtagg agccgccggc gaagatccag gaccaggacc cgcaccaccc
     4201 cccgaaggag ctcaggaagc tgcccccgca cccaggccac ctcctgagcc cgacgatgcc
     4261 gcagctgctc tgcgacttgc attggaccag ctttccgctc tcggactggg gggagcaggc
     4321 gataccgacg aggagggggc cgctggcgat ggcgccgcag cagctggcgg agctgatgga
     4381 ggggcagcac ccgagcccgt tcctccagac ggaccagagg caggggctcc acctacactc
     4441 gcacctgctg tcgcccccgg atcacttcct ttgctcgatc ccaacgcatc cccacctcct
     4501 ccaccacccc cacggccttc acctcctgat gtgttcgcag gatttgctcc tcatcccgct
     4561 gcccttggcc cacccactct gctggctgat cagatgagcg tcattggtag tcggaaaaaa
     4621 agtgtgaata tgaccgagtg cgtcccagta cccagttctg aacatgtggc tgaaatcgtt
     4681 ggccgacaag gacccagaac cgtcatccat ccaaaagctc ggataatagc cgaggcaggc
     4741 cccatagtca ttggagaagg gaacttgatt gaggaacagg ctctgataat taacgcttat
     4801 cctgacaaca tcacccccga caccgaagac ccagagccta agcctatgat tatcggtact
     4861 aataatgtat ttgaggtagg atgctactca caggcaatga aaatgggaga taacaacgtt
     4921 attgagtcta aggcttacgt tggcaggaac gtcatcctca cttccggctg cattataggc
     4981 gcctgctgta atttgaacac tttcgaagta atccccgaaa acacagtgat ctacggagcc
     5041 gactgcttga gacgcgtcca aactgagcga cctcagcctc agacactcca attggatttc
     5101 ctgatgaaga ttctgccaaa ctaccaccac cttaaaaaga ctatgaaagg aagttccaca
     5161 cccgtaaaga attgaaccgg ttctagataa gattacggta cctttaagac caatgactta
     5221 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5281 tcactcccaa agaagacaag atctgctttt tgcctgtact gggtctctct ggttagacca
     5341 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     5401 cttcagctgc tcgagctagc agatcttttt ccctctgcca aaaattatgg ggacatcatg
     5461 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     5521 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     5581 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     5641 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     5701 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     5761 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     5821 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     5881 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5941 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6001 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6061 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6121 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6181 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6241 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     6301 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6361 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     6421 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6841 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6961 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7021 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7081 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7141 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7201 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7261 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7321 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7381 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7441 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7501 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7561 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7621 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7681 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7741 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7801 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7861 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7921 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7981 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8041 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8101 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacc
//
