LOCUS       sequence-407473-        8547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, SS18_SSX1 from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8547)
  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 8547)
  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_SS18_SSX1
FEATURES             Location/Qualifiers
     source          1..8547
                     /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(3308..4018,4019..4021,4022..4024))
                     /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(3350..3371)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3699..3718
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3958..3979
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4021..4030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4106..4309)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4135..4155)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4310..4613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4870..4914)
                     /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    5099..5332
                     /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    5692..5809
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6301..6426
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6473..6730
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6732..7111
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7142..7246
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7247..7315,7316..8107)
                     /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(7465..7484)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8278..8547,1..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 ccggttcatt
     1801 catcatcttc ctctggatca gaaatctctt catagattac taattgttta cgttcacgaa
     1861 ggcgatgtgt ccacgcatgc ttgccacgtt ttgggcctga acgcttatta attttttcgc
     1921 taatatttgc tttaccagga ggatgtaact gtttaccatc attctgtggg ccactagcct
     1981 ctgagactcc tttagagtcg ttttcgtctt ctgccggctt ttttggcatg atctggtcgt
     2041 acccgtatgg gcgctgctgc ggtggttgag gtggcccagg ttgcgtggga cggtatccac
     2101 catactgctg gccttgtcct tgaggatagt tggggtattg cggtcctggt ccaccttgtg
     2161 acggtccgta gccctgctgt tgacctgggt agccttgttg acccggatat tgctgttgct
     2221 ggggaggata tccctgttga ggcggaggcc cttggtaggc gtcttgctgc tgcccatatt
     2281 gggagtttcc tccctcatag tagtgctggg aactatcttc atagggacga tcgtagccct
     2341 gttcggggta cgaaggttgc tggtagccat agtcgttgtg accgtctgga taatactgtt
     2401 ggttcatccc ttccggcggc ccctgacccc cgtgactata ttgatctcca tagtagtcct
     2461 cttggcctga atactgttgt ggcggtccct gttggggagg gcgataagga gggatctggc
     2521 gttggcccat catgtgattg ccctggttca cttgccccat catccccatg ggaggctgct
     2581 gaccttgata atgttgtcca ccaccctgtg gcatgttgta ctgttgggag ggaggttgct
     2641 gatgcatcat cggaccctga tttggttgca tggacatatt cgggcgtggc ccgtaattac
     2701 ccataggctg gccctgcgac attgtcattt ggttttgaac cggcatagac tgtgaggatg
     2761 gtaccgagtg gttgtagcca cccatggagc catgcgagct ggaaggcata ttcatactcg
     2821 aattagtcat gttaagttgg ttaggaccag gtccctgcat gggcatgtgg ttggggccgg
     2881 gcatctgacc gttcatctgg ttctgcatgt gcggtgctgg gggtccgcca ccgaccatcc
     2941 catcggatgg catgttatgt gagcgagggg gaggtggtgg acccgactga ttcatgcccc
     3001 cagggcccat gggcatattt tgcgtaggtg gcgctggcaa caacgattgc atgttctggt
     3061 tgctgtctgc gatagtagcc aagtaaacaa gattcgtatg caacatctgt tgatattgag
     3121 aacattccga ggttttgccc ttgttttgac tatccatgat gcattggatt aagtggttat
     3181 tgtcatcaag catcttttgg atcgcagcgg gcgtaatctc gccttttcca cgctgacgcg
     3241 gggccgcgaa cgcgactgac attccccggg aagcggccgc acttcccccg gagcctccgc
     3301 ttccgccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     3361 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca
     3421 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     3481 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     3541 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     3601 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     3661 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     3721 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     3781 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     3841 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     3901 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     3961 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     4021 ccatggtggc gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcggttca
     4081 ctaaacgagc ccggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc
     4141 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     4201 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     4261 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     4321 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4381 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4441 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4501 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4561 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4621 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagcacgcg
     4681 ttcgaagctt gagctcggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga
     4741 atatccctgc ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac
     4801 caagcctcct actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa
     4861 ccaattccac aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg
     4921 ctggttttgc gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct
     4981 gtcccactcc atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact
     5041 agcattccaa ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag
     5101 gagctgttga tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat
     5161 gccccagact gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt
     5221 ctgctgctgc actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc
     5281 tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac
     5341 tgctcttttt tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg
     5401 ttcaattttt actactttat atttatataa ttcacttctc caattgtccc tcatatctcc
     5461 tccttatggt gttttactaa acttttccat gttctaatcc tcatcctgtc tacttgccac
     5521 acaatcatca cctgccatct gttttccata atccctaatg atctttgctt ttcttcttgg
     5581 cactactttt atgtcactat tatcttgtat tactactgcc ccttcacctt tccagaggag
     5641 ctttgctggt cctttccaaa gtggatttct gctgtccctg taataaaccc gaaaattttg
     5701 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
     5761 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
     5821 tactgccatt tgtactgctg tcttaagatg ttcagcctga tctcttacct gtcctataat
     5881 tttctttaat tctttattca tagattctac tactccttga ctttggggat tgtagggaat
     5941 ttccaggtct gaagatcctg ctgcttgctg tgcttttttc ttacttttgt tttgctcttc
     6001 ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata
     6061 gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg
     6121 ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc cagtattaac
     6181 tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt
     6241 tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact
     6301 gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact
     6361 caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg
     6421 agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag
     6481 attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga
     6541 cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc
     6601 cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag
     6661 gcaaaaagca gctgcttata tgcaggatct gagggctcgc cactccccag tcccgcccag
     6721 gccacgcctc ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     6781 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     6841 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     6901 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     6961 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     7021 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     7081 tactattaat aactagtcaa taatcaatgt cgacccaggt ggcacttttc ggggaaatgt
     7141 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7201 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7261 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7321 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7381 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7441 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7501 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7561 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7621 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7681 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7741 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7801 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7861 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7921 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7981 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8041 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8101 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8161 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8221 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8281 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8341 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8401 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8461 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8521 cagtggcgat aagtcgtgtc ttaccgg
//
