LOCUS       sequence-406596-        8622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, BTN3A3_BTN3A1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8622)
  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 8622)
  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_BTN3A3_BTN3A1
FEATURES             Location/Qualifiers
     source          1..8622
                     /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(3383..4093,4094..4096,4097..4099))
                     /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(3425..3446)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3774..3793
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4033..4054
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4096..4105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4181..4384)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        4385..4688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4945..4989)
                     /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    5174..5407
                     /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    5767..5884
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6376..6501
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6548..6805
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6807..7186
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7217..7321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7322..7390,7391..8182)
                     /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(7540..7559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8353..8622,1..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 ccggttcacg
     1801 cagggcaaat agtcagtgca gtcggttcca aggttaaaat acggaagacc ggataaagtg
     1861 cctcggaaaa actcacatca aggaatgtat gaatgtgcga accatccacg gcgttgtaaa
     1921 acgagatatc ccctgtctcg tagtccaaga acacgcctac cttcttgggc ggttttggca
     1981 gtttaagatt cgtacggggt tctgtcagcg tgcgatactt attaccatca gttaacccca
     2041 tagtccagaa tccattctca ggagtcatct tgacccaacc tttgcgctgc acatttttag
     2101 aacatacacc gatatgccac tctttgcggt caccgacctc cacttcccaa tagtggcggc
     2161 cactaatgaa actctcacac cctaatacgc aataatgcca attgaaacgc tccggattgt
     2221 ctggcaggtc ttgaggttct ttcgcacgtt gtaccgaacg ttgatcctct gaaaccaaaa
     2281 gaatagggtt agcagtcttc gggtccagga tcacatccgc cggcttaaac aacgccatct
     2341 tccattcgtg ataggcaaga gatttctcgc cacgagccat gtattgaatt ttgcgccatt
     2401 tcagttcttc ctgcagtttt tcacgcaggc tgatctcctg ttcagtcgct gcatagccca
     2461 tctctttcat ctcgcgttca cgttctgtct cgcgtgacag cgcgatcttt tctttctgct
     2521 ggcgccacaa aaagtaagac gcccccgcaa gcagcagcaa gctaattggt aaagttcctg
     2581 ccaatgctgc gatccaaggt tgggcggaac gaaagaaagg gtctgcaatg gagattgaag
     2641 cagttttttc caaacccaga agagagttac gaataatgca ggatacgccg ccaccagagc
     2701 ttccacgcat aataactgac gcggcgacag cataaaggcc tacaccgtct gcgaccactg
     2761 gtgcttctac cgctggaatg ttctcgcctt tggtatctga ccacttaatt tgtggctgag
     2821 ggtaccaccc cgtactgcga cattccaaat gaatgccacc atcctcatat ccctttactt
     2881 cgatatgaag gtcgctcccc agagcggcta ccttcagctc gacaagcgcc ttctcgtaga
     2941 agtcgccgtc ctgaaaataa cataaatatt tccccgagtc actcgcagtg acattgtgaa
     3001 tacgcaaggc ggctttaccc gccgtgatgc catcacgcag gatgcttgta cgtccgcggt
     3061 acggagcaga ttggcggtcc tctacctctt tgccatccgc gtaaacattt acgacttggc
     3121 gcagggacga cgagacccag cgtaattcca tagtctctgc ggacatggtt ggaaaaaggt
     3181 gacaaggtaa gtccgcatcc tcgcccacca tagccagaat gggccccgag gggcccagca
     3241 cagaaaactg cgccgagcac ggagtaagta actgtaccaa aaacaggctc acatgaaaat
     3301 tcagaagcaa gaacgctaat gaagatgcca ttttcattcc ccgggaagcg gccgcacttc
     3361 ccccggagcc tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg
     3421 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct
     3481 tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     3541 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     3601 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     3661 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     3721 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     3781 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     3841 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     3901 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     3961 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     4021 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     4081 cctcgccctt gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact
     4141 gcagaattcg gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct
     4201 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     4261 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     4321 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     4381 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     4441 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     4501 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     4561 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     4621 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     4681 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     4741 aataactagc acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg
     4801 aaacgataat ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac
     4861 tattcttaaa cctaccaagc ctcctactat cattatgaat aattttatat accacagcca
     4921 atttgttatg ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg
     4981 ttcattcttt tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta
     5041 attgttaatt tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga
     5101 tttattactc caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca
     5161 accccaaatc cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc
     5221 ctggagctgc ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc
     5281 cctcagcaaa ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt
     5341 cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa
     5401 agctcctatt cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg
     5461 tgctactcct aatggttcaa tttttactac tttatattta tataattcac ttctccaatt
     5521 gtccctcata tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc
     5581 ctgtctactt gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt
     5641 tgcttttctt cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc
     5701 acctttccag aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata
     5761 aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt
     5821 tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt
     5881 aaaattgtgg atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct
     5941 tacctgtcct ataattttct ttaattcttt attcatagat tctactactc cttgactttg
     6001 gggattgtag ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact
     6061 tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct
     6121 ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc
     6181 tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa
     6241 caggccagta ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa
     6301 tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct
     6361 cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa
     6421 aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag
     6481 ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg
     6541 gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca
     6601 gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc
     6661 ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag
     6721 agagacccag tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc
     6781 cccagtcccg cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac
     6841 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     6901 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     6961 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     7021 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     7081 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     7141 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac
     7201 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7261 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7321 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7381 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7441 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7501 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7561 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7621 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7681 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7741 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7801 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7861 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7921 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7981 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     8041 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8101 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8161 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8221 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8281 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8341 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8401 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8461 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     8521 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8581 accagtggct gctgccagtg gcgataagtc gtgtcttacc gg
//
