LOCUS       sequence-408182-        7662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, ZFAND3_DGKH from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7662)
  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 7662)
  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_ZFAND3_DGKH
FEATURES             Location/Qualifiers
     source          1..7662
                     /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(2423..3133,3134..3136,3137..3139))
                     /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(2465..2486)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2814..2833
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3073..3094
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3136..3145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3221..3424)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3250..3270)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3425..3728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3985..4029)
                     /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    4214..4447
                     /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    4807..4924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5416..5541
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5588..5845
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5847..6226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6257..6361
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6362..6430,6431..7222)
                     /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(6580..6599)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7393..7662,1..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 ccggttcaat
     1801 tcccaggact gttcgcttca caggaaccgg attccgtatc tccactccca caacggcatg
     1861 aacctaattc ttgctgcact aattccagct tagtttgaca ctgaaaacaa cggcggcggg
     1921 acttttgctt cgagcgactg gtctcctcgc tacgttcggt gttctccagt aaacgggggc
     1981 gttttacggg gctcgcttca ttttccgatt ggctgtccgt gccgcacgag cgtttggtgg
     2041 gcgtgataag ggatacgtgg gctgtgtccg tgcacggccc gcattcttct ttgcttggac
     2101 tcgtgacgtt aagctctgta ggtaagggct gttgacttgg agacaaagta ggtgtggtga
     2161 tggacgtgtt attgttatca ctagtagttt cctcgctgaa aaggtcggac tgcgaattgg
     2221 aagttgaagg tgcgctgtca tcgtccggtt gcttcttctg aaaatccgca aaacatttgg
     2281 aacaaagatt catggtcttg ctagaacccc aaaagccaca cgggcagcga ggaggcagcg
     2341 agggggcctt actacgctca gagcctgcgt cacccattcc ccgggaagcg gccgcacttc
     2401 ccccggagcc tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg
     2461 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct
     2521 tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     2581 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     2641 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     2701 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     2761 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     2821 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     2881 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     2941 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     3001 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     3061 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     3121 cctcgccctt gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact
     3181 gcagaattcg gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct
     3241 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     3301 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     3361 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     3421 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     3481 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     3541 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     3601 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     3661 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     3721 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     3781 aataactagc acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg
     3841 aaacgataat ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac
     3901 tattcttaaa cctaccaagc ctcctactat cattatgaat aattttatat accacagcca
     3961 atttgttatg ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg
     4021 ttcattcttt tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta
     4081 attgttaatt tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga
     4141 tttattactc caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca
     4201 accccaaatc cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc
     4261 ctggagctgc ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc
     4321 cctcagcaaa ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt
     4381 cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa
     4441 agctcctatt cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg
     4501 tgctactcct aatggttcaa tttttactac tttatattta tataattcac ttctccaatt
     4561 gtccctcata tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc
     4621 ctgtctactt gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt
     4681 tgcttttctt cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc
     4741 acctttccag aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata
     4801 aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt
     4861 tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt
     4921 aaaattgtgg atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct
     4981 tacctgtcct ataattttct ttaattcttt attcatagat tctactactc cttgactttg
     5041 gggattgtag ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact
     5101 tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct
     5161 ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc
     5221 tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa
     5281 caggccagta ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa
     5341 tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct
     5401 cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa
     5461 aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag
     5521 ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg
     5581 gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca
     5641 gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc
     5701 ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag
     5761 agagacccag tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc
     5821 cccagtcccg cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac
     5881 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     5941 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     6001 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     6061 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     6121 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     6181 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac
     6241 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6301 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6361 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6421 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6481 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6541 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6601 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6661 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6721 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6781 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6841 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6901 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6961 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7021 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7081 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7141 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7201 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7261 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7321 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7501 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7561 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gg
//
