LOCUS       sequence-407455-        8094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, RANBP3_SLC25A23 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8094)
  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 8094)
  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_RANBP3_SLC25A23
FEATURES             Location/Qualifiers
     source          1..8094
                     /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(2855..3565,3566..3568,3569..3571))
                     /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(2897..2918)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3246..3265
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3505..3526
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3568..3577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3653..3856)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3682..3702)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3857..4160
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4417..4461)
                     /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    4646..4879
                     /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    5239..5356
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5848..5973
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6020..6277
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6279..6658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6689..6793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6794..6862,6863..7654)
                     /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(7012..7031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7825..8094,1..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 ccggttcaac
     1801 gtgatgtgac gcctaatgcc tgtttcatgt tctcgtacac tacgtacgaa atgctcacgg
     1861 cgggaatcac cttcataaaa tttggcgcaa ttccacgata aagtccgcgc attccctcct
     1921 gcgataagat atggcgaagc aagcccagca tagacaactg cggaccgcct tcaatactgg
     1981 cttgggcctg catgcgggtg cgtacaagag ccagcgggta cgaggcaatc tggccgcaag
     2041 tagatgagat agtcccacaa gcaagcaaga ccagaatacc tggatctgca gagtcatgac
     2101 tgtactgttg cagccaccag ttcttaaggg tctcatctgc ggcctccgag gggctgcgcc
     2161 aagccgccgt gtcgggactt gcagccggaa cggcccccgt acaatctgct ggcaaagaca
     2221 cgccatttgt gccgctgctt ggaaccgttt gagacagtgc cttaggttgc ggggcttgca
     2281 gtacggctgg acgcaggact gagcgttgct gctccttcgg tttttgcgat ggtaaccctg
     2341 cagaaggcgc ttggccgtgg ataagggttg ggggctttaa acgaaacccg gaactgcgtt
     2401 cctctgattg actagggggg aactgcgtca atgaggatgt gcgttcgcgt ttaacaggtg
     2461 gacaataatt tccgtcttca cggtccgaat cctccccacc ctcgggagaa ctaccgcctg
     2521 cgctgcgccc agctaactca cgggggaaag gaggaagttg agcctctggc gcagggggtg
     2581 gaggtgttga ggctgatgct ccagccggag caggtggttc cagggcgtgc tcccccgcag
     2641 attccgggtg tccagtaccg tgatgtggag cctcagcctc accacgtggc tcctctccac
     2701 tatcggacag atttttttgt tcggcgggac tcttttgacc cttatctttc tgaaacacga
     2761 aaaccggggg cgcgattgca ggtttttctt cgttggccaa gtctgccatt ccccgggaag
     2821 cggccgcact tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag
     2881 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     2941 ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     3001 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     3061 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     3121 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     3181 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     3241 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     3301 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     3361 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     3421 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     3481 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     3541 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg
     3601 gtaccgtcga ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc
     3661 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     3721 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     3781 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     3841 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     3901 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     3961 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     4021 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     4081 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     4141 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     4201 ttgattacta ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg
     4261 gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag
     4321 tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga ataattttat
     4381 ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc
     4441 caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat
     4501 taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat
     4561 ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag
     4621 ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc
     4681 caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg
     4741 cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct
     4801 ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa
     4861 cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt
     4921 tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt tatataattc
     4981 acttctccaa ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt
     5041 ctaatcctca tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc
     5101 cctaatgatc tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac
     5161 tactgcccct tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct
     5221 gtccctgtaa taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt
     5281 ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc
     5341 cccttttctt ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc
     5401 agcctgatct cttacctgtc ctataatttt ctttaattct ttattcatag attctactac
     5461 tccttgactt tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc
     5521 ttttttctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt
     5581 tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag
     5641 ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc
     5701 tgatgtttct aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac
     5761 tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg
     5821 cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc
     5881 tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg
     5941 cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa
     6001 gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat
     6061 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
     6121 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
     6181 tggtctaacc agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag
     6241 ggctcgccac tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat
     6301 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     6361 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     6421 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     6481 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     6541 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     6601 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga
     6661 cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6721 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6781 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6841 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6901 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6961 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7021 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7081 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7141 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7201 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7261 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7321 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7381 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7441 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7501 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7561 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7621 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7681 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7741 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7801 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7861 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7921 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7981 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8041 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgg
//
