LOCUS       sequence-407538-        8415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, LMX1B_GTF3C5 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8415)
  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 8415)
  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_LMX1B_GTF3C5
FEATURES             Location/Qualifiers
     source          1..8415
                     /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(3176..3886,3887..3889,3890..3892))
                     /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(3218..3239)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3567..3586
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3826..3847
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3889..3898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3974..4177)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4003..4023)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4178..4481
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4738..4782)
                     /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    4967..5200
                     /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    5560..5677
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6169..6294
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6341..6598
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6600..6979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7010..7114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7115..7183,7184..7975)
                     /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(7333..7352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8146..8415,1..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 ccggttcaaa
     1801 cataatcaag aatctccgtc tccatctcgt tctcggaacc atcgctgggt ttaaagtcct
     1861 cttcttcttc ttcctcctct tcctcgtcct cttcatcttc cccgctttca taggtgagct
     1921 gctccttccc gccgtcagcc ttggcactac ttgaaaagag tgcaggtcgc tttgagcgga
     1981 tggtttgccg aatcatcaat gacatcgtat ctcgcaactc atcgctcgtt ttggggagac
     2041 accacccatc gcgttctgta catgaatttt ctgccccatc atttcgatgt atgatcttct
     2101 ggagttcttc tacattcaag tcacagagtt gataaaacat ctgtcgataa ggaggaagtg
     2161 cgccttcgcg aaatatatac acggagtccc taagtgtctg caaggagact ttcagcttat
     2221 atttagaaga agctggcttc ctcgcaccgg acgtaccaga ggggcccaag ccctgtttaa
     2281 ggtcatgcat cgtcaccaac tgtgagctag tcttcttcac tgttatgggc aaactgtaat
     2341 tgtaagtgct tcgcttggcc ttcacaggga ggtctgaagg agcgtatcca tgcttcatcc
     2401 cacaccgtat tcggaaatca agaacctggt agatcttggc gtcggggttc ttcctcgggt
     2461 cgtagccgaa gcgtatccac aggcttctcc atggccctgt gatcatatag taagcgataa
     2521 aagggaggag aaccttaagt ttgtctgggt gaacggatat attcgctttc actgcgttcc
     2581 tactccagat tggtcggatg tcgaagagtt tccgaagttc ttcttccact ttcctgtcca
     2641 ctggatttgt gcatacccgt cgccatgtct gagctgcagc ttccaaaggt tgtttcggga
     2701 cttcttcgtc ctcgaagttt acaaagatgg cattgtgagg gcgtcgagcg cgtgacaaac
     2761 ctatgagatt ttcaccactg ataggggggt tgttgtaacc ttcttgctgg tagtcttgtt
     2821 tgcaataaag ttttcggtcc cgaaagtaac aagacgttgt caaggcttgt tgacaagcgg
     2881 cacactgcag gcactcttca tgccaggaac tctcgttaac tctcatcaaa aagcggtcgc
     2941 tgatgggccg ctggcacccc tcgcagaccg cagggtgtgg acaatctgat ccaagaagca
     3001 cgcccaacgt tgctggccct ggccgcagag catgctcttc cattttaatc ccgtcaagca
     3061 tttttgcgca atctgtttga ccccttggga agcagcgttc cagactttcc gggcctgtgg
     3121 ctatatccat tccccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt
     3181 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     3241 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     3301 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     3361 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     3421 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     3481 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     3541 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     3601 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     3661 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     3721 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     3781 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     3841 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     3901 ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc
     3961 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     4021 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     4081 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     4141 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     4201 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4261 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4321 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4381 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4441 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4501 tatgaactaa tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga
     4561 gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
     4621 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
     4681 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
     4741 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
     4801 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
     4861 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
     4921 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
     4981 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
     5041 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
     5101 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
     5161 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
     5221 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
     5281 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt
     5341 tttactaaac ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc
     5401 tgccatctgt tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat
     5461 gtcactatta tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc
     5521 tttccaaagt ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     5581 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     5641 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     5701 tactgctgtc ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc
     5761 tttattcata gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga
     5821 agatcctgct gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc
     5881 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     5941 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     6001 cagtatttgt ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc
     6061 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     6121 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     6181 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     6241 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     6301 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     6361 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     6421 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     6481 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     6541 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     6601 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6661 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6721 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     6781 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6841 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6901 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     6961 ctagtcaata atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7621 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7801 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7921 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7981 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8281 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8401 gtcgtgtctt accgg
//
