LOCUS       sequence-407531-        8721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MFF_ERBB4 from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8721)
  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 8721)
  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_MFF_ERBB4
FEATURES             Location/Qualifiers
     source          1..8721
                     /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(3482..4192,4193..4195,4196..4198))
                     /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(3524..3545)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3873..3892
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4132..4153
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4195..4204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4280..4483)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4309..4329)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4484..4787
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5044..5088)
                     /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    5273..5506
                     /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    5866..5983
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6475..6600
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6647..6904
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6906..7285
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7316..7420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7421..7489,7490..8281)
                     /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(7639..7658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8452..8721,1..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 ccggttcaaa
     1801 caactgtgtt gcggtggcga tacggtggtg gtggcagaac ggtccctggt ttcaacgaga
     1861 actctgataa atactctgga ttttccgcca cgatcgggcg aatacggccg ttctgtttat
     1921 aaaagtactt agtggagtat tcttgcaagt aatctgggtg ttgaagggtg ctacgagggg
     1981 gcaaactgtg gttccagtag tctggattat caaaggcttt cttggccttc tcaggcatgc
     2041 tcaaaatatt gttttttaag tactccgcct tacctaaagt gttcgcgaag gtgttcaggt
     2101 aaaggggctc gttgacgtac tcatcttccg ccttaggcgg cccgttagac gcattgtggt
     2161 attctggatt gtctaacgcc tgcagatccc catttttacg acgggatacg aagggatttt
     2221 cctctacagg attcaggtat tcttgtttcg gcttgtcacg cattggtgtc atgtagccct
     2281 cctcgtccag ctccccacgc ggactacgct ccggtgcgaa aaccgtggga tctgcactgt
     2341 aacgctgtgt gctagaatct tcctgaacgt gaggcgcgac tggcttgcgc aaagtcccat
     2401 tgcagcaaga atcatcaaaa atctcagctg tggccccttg tgcaactggc gcctcaggga
     2461 tagtactagt aggagcgcgg tagggcactg agactccctg ttccgccgcg aaccccccat
     2521 cacggtagac aaattggtta cctgacatgg gagtataggc gggcggaggc gagtgcccaa
     2581 tttccgagcg attggaatca atgcgcgcgc gactagtata gataggtggc gggatattga
     2641 atgcctgcgg gacaaggtac tcttcagcat ccatcatgtc ctcaagatct tcctcatcaa
     2701 gcaggttttg gaaaaacttg ctatcgttgg gactaggcag cttcatgcga tcgtcaccct
     2761 gaataactaa gtaacgctga ggatcacggg ccatacgtga aaattctgcg gccaattctt
     2821 tgaactttgg gcgagaatcg gcgtcgatca tccaacattt gaccataacc atgtaaacat
     2881 cgattgtgca gatcggaggt tgtggtaaac gctcgccttt ctccaacagg tccgggattt
     2941 cgcgcgtggg gatgccgtca tacggcttcc ctccgaaagt cataagttcc caaatcgtaa
     3001 ctccgtatga ccaaacgtca ctttggtgag taaatttacg atagtggatg cactccagtg
     3061 ccatccattt gatgggcatc ttcccaccat ctgcgttgta ttccttttca tcgccctcca
     3121 gtaaacgggc cagtccgaag tcagtaattt tgacatggtt cggggattta acaagcacgt
     3181 tacgcgctgc aaggtcacga tgtacaaggc ggcgctcttc caagtacatc attccctcct
     3241 cgttttgcgg tgtggtcggg ggacgttcta agtccaaaaa atccaaagga cgctcagata
     3301 aagtcagaac gcgcggggga gttttaagcg ccaagggttt aaaaggagtt gactggatta
     3361 aatccaagtc cgctgggcga gaaaaggata cgtcctcatt gttgcctgca acaacgatac
     3421 gttccggcac ctgcattccc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc
     3481 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     3541 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt
     3601 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     3661 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     3721 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3781 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3841 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3901 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3961 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     4021 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     4081 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     4141 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     4201 tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac
     4261 gagcccggtt cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     4321 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     4381 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     4441 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     4501 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     4561 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     4621 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     4681 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     4741 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     4801 atgggctatg aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa
     4861 gcttgagctc ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc
     4921 ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc
     4981 tcctactatc attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt
     5041 ccacaaactt gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt
     5101 ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca
     5161 ctccatccag gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt
     5221 ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg
     5281 ttgatccttt aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca
     5341 gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg
     5401 ctgcactata ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc
     5461 catagtgctt cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct
     5521 tttttctctc tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat
     5581 ttttactact ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta
     5641 tggtgtttta ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc
     5701 atcacctgcc atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac
     5761 ttttatgtca ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc
     5821 tggtcctttc caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt
     5881 tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct
     5941 ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc
     6001 catttgtact gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt
     6061 taattcttta ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag
     6121 gtctgaagat cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat
     6181 cttgtctaaa gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt
     6241 tgctactgta ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag
     6301 ctgtcccagt atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa
     6361 tcgttctagc tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc
     6421 tggccttaac cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct
     6481 ctcgcaccca tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag
     6541 tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag
     6601 ctcctctggt ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc
     6661 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     6721 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     6781 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa
     6841 agcagctgct tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg
     6901 cctcccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     6961 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     7021 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     7081 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     7141 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     7201 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     7261 taataactag tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg
     7321 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7381 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7441 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7501 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7561 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7621 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7681 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7741 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7801 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7861 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7921 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7981 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8041 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8101 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8161 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8221 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8281 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8341 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8401 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     8461 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     8521 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     8581 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     8641 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     8701 cgataagtcg tgtcttaccg g
//
