LOCUS       sequence-407471-        8304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, SPAG9_SKAP1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8304)
  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 8304)
  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_SPAG9_SKAP1
FEATURES             Location/Qualifiers
     source          1..8304
                     /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(3065..3775,3776..3778,3779..3781))
                     /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(3107..3128)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3456..3475
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3715..3736
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3778..3787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3863..4066)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3892..3912)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4067..4370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4627..4671)
                     /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    4856..5089
                     /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    5449..5566
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6058..6183
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6230..6487
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6489..6868
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6899..7003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7004..7072,7073..7864)
                     /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(7222..7241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8035..8304,1..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 gttcctcgac ctcaaaggca gtagttaagt attccttagg gacaataccc accaatgaat
     1861 ttaactcacc gacccaccaa ccatacatgt tgtactcttt cgacaggata cggataagat
     1921 ccccacgttg gaagctcagt tcgtcaggct ggtcgccgtg gcaatcccac aatccttgat
     1981 aatacgacgc atagtcaacc cccttgcgac gtgtgccaga ctcatcttct tcaagatcgt
     2041 gctcctcgtc aggtaagact tcatagatgt cttcctcctc cttctcttct gttggctctt
     2101 taatcccaac tgaacccggc aagatagtgg gacgacattg acttccgcac gagggagagt
     2161 cgaacccatc gatgtcgtca tacgtctcct ctttctcctc ctcctcctcg tcctcttcat
     2221 aagggatggt cagggaactc aaatctttca aaaggaaact gatctggtca acccagtcac
     2281 gcgcttctgc tgggctagtg gcggtgaact catagctacg gcgatcttgt gatgtaagtt
     2341 cgaagcaact ttctttttta ctatcacggc gtaagtgggg agccatacgg acaccatacc
     2401 ccttaatcaa gaaggtgcct ttcggttgtt tagacttttc attagcgtag taataaaaca
     2461 gaccgcgcga cacaacgcac caacgttttt gccattcact gccaaaaaag ctatggtctt
     2521 tgctcttttt ttctaagtag ccctgcttga tgacgttgtc caactcctgg gctcctttta
     2581 cgatgtcttc catgcgctgg tcggcgtaat tcttggcctt caactctaac tgacgtgtct
     2641 ggctctcaag actttccacg cgcgtctgca ggtccttctt ctcctgctcc tgggaatctt
     2701 caaattcgat gaatttctcc tcagcgtgtt tgcgtaacgc cttctcacgt tcatactgcg
     2761 taataagttg ctcgttgtca tcacgtaaca attccaattc tacctggtgt tcctggtcct
     2821 gtgcaaaaac ggagtccagg ttttctaaaa ctgcgacaac cagcggcatc agttccttca
     2881 ctacttcttc atcatagcga ccgataagac gctcgaactc gcggtaaata cttcccgcca
     2941 ggccgctcac acgctctgac atgactgcgc cagaacctcc gggctcttct tggtagacca
     3001 cgccgtcttc taactccatt ccccgggaag cggccgcact tcccccggag cctccgcttc
     3061 cgcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     3121 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg gtgctcaggt
     3181 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     3241 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc
     3301 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     3361 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     3421 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     3481 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     3541 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     3601 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     3661 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     3721 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     3781 tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga ctgcagaatt cggttcacta
     3841 aacgagcccg gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac
     3901 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     3961 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     4021 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     4081 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4141 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4201 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4261 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4321 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4381 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gcacgcgttc
     4441 gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
     4501 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
     4561 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
     4621 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
     4681 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
     4741 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
     4801 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
     4861 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     4921 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     4981 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     5041 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     5101 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     5161 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     5221 ttatggtgtt ttactaaact tttccatgtt ctaatcctca tcctgtctac ttgccacaca
     5281 atcatcacct gccatctgtt ttccataatc cctaatgatc tttgcttttc ttcttggcac
     5341 tacttttatg tcactattat cttgtattac tactgcccct tcacctttcc agaggagctt
     5401 tgctggtcct ttccaaagtg gatttctgct gtccctgtaa taaacccgaa aattttgaat
     5461 ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat
     5521 tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt ggatgaatac
     5581 tgccatttgt actgctgtct taagatgttc agcctgatct cttacctgtc ctataatttt
     5641 ctttaattct ttattcatag attctactac tccttgactt tggggattgt agggaatttc
     5701 caggtctgaa gatcctgctg cttgctgtgc ttttttctta cttttgtttt gctcttcctc
     5761 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag
     5821 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg
     5881 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag tattaactgc
     5941 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc
     6001 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac
     6061 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac
     6121 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga
     6181 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt
     6241 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     6301 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     6361 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca
     6421 aaaagcagct gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc
     6481 acgcctccca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6541 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6601 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     6661 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     6721 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     6781 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     6841 tattaataac tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg
     6901 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6961 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7021 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7081 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7141 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7201 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7261 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7321 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7381 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7441 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7501 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7561 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7621 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7681 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7741 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7801 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7861 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7921 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7981 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8041 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8101 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8161 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8221 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8281 tggcgataag tcgtgtctta ccgg
//
