LOCUS       sequence-406604-        8592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, CCDC6_RET from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8592)
  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 8592)
  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_CCDC6_RET
FEATURES             Location/Qualifiers
     source          1..8592
                     /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(3353..4063,4064..4066,4067..4069))
                     /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(3395..3416)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3744..3763
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4003..4024
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4066..4075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4151..4354)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4180..4200)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4355..4658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4915..4959)
                     /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    5144..5377
                     /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    5737..5854
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6346..6471
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6518..6775
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6777..7156
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7187..7291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7292..7360,7361..8152)
                     /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(7510..7529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8323..8592,1..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 ccggttcagc
     1801 tgtcaaaagt gtccattaat tttgctgcgg acggactcaa catccaatta gcgtagacgc
     1861 tgtcattagg ataacgggga aaaccagtat ttgtgccatc tgcacgcgta agtggaacag
     1921 gagactcacc cggccagtta ggatcactca tcccatacaa cttattttcg atccaggttg
     1981 agggcagggc gcggggcaag ggagcattgt tgcagtctac cagtggcgtt tcttcctcgg
     2041 ataagccgtc atcgtagata aggctatcac ttggcgtgct ggctgctaaa tccaaataat
     2101 cacgacgttt caccatcatt ttttccaaat cctttgagat gtcagcgaag acaggacgtt
     2161 tgtcgggctc ctgtttccaa cactgcaaca ttaagcgata catttcttca gagcaattat
     2221 cggggcgctc catacggtgt cctgttttta aaaggttgaa caaacgctcg ggcggaatac
     2281 ctgggtaggg attgccacct aaagtcacaa tctcccaaag caagacaccg aatgaccaga
     2341 cgtcactctg cgttgtgtag atgtgatcaa acaaactctc aatagccatc cacttgactg
     2401 ggatacgtcc ctgactacgc ttaacataac tatcttcctc gtatacgtca cgtgacagtc
     2461 caaaatcgct gatcttcatt ttacgccctt cagctacaag gatattacgc gctgcaagat
     2521 cgcggtgtac cagcttcatt tctgccaggt attgcatacc ctgtgaaatc tgccacgcaa
     2581 aggaaatcag atcccccatt gtcaatgcgc gttcatccgg gtgatctaat gaactcgaat
     2641 tacgggaccc gcctgatccc aagtaaccag gcccaacctt acgagattcg cgtaagaacc
     2701 cgcgaagact gccgtatttg gcgtactcga cgattaacag aagcggtcca tcctgactac
     2761 atgcgccata taatttaatc acatgtggat ggttcacttg tttaagtaca ttaaactcgg
     2821 acagcaggtc acgcaattca gacgggctgg cgttttcttt caacattttt accgcgacag
     2881 ttgtatagcc tgcgcgacct ttaaggtgaa aagctgtagc tttcacgact ttgccaaact
     2941 ccccttcccc caatgtctta cctaacacaa ggtttttgcg aggaaattcc catttcggat
     3001 cttcgatcgt tacggacgcc ttacgaagat cacgattctc ctcttgcaga gccttacatt
     3061 ttaatttgta ggtttccaat tcgatcttca gcactttgtt ttcttgttgc agactggcca
     3121 agcgattagt aagctcttcc aggcgaaagg gtgagataac aatgcctcca ctcttaccgc
     3181 ctccgcctcc tccgcccccg cctccaccgc cccctgaagt gctagagcaa gacgattgca
     3241 tagcagcgga tgaactagag ttacctcccg ccccatcggt gtcactctcc gaggcgctgt
     3301 ctgccattcc ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca
     3361 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     3421 cgcgcttctc gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg
     3481 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     3541 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     3601 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     3661 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     3721 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     3781 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     3841 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     3901 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     3961 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     4021 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg
     4081 gtggatcccg ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt
     4141 tcactaaacg agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     4201 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     4261 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     4321 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     4381 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4441 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4501 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     4561 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     4621 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     4681 gaactaatga ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct
     4741 cggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac
     4801 tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat
     4861 cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact
     4921 tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc
     4981 ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca
     5041 ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac
     5101 agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt
     5161 taggtatctt tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag
     5221 ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat
     5281 accagacaat aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct
     5341 tcctgctgct cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct
     5401 ctgcaccact cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac
     5461 tttatattta tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt
     5521 actaaacttt tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc
     5581 catctgtttt ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc
     5641 actattatct tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt
     5701 ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg
     5761 tttttgtaat tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc
     5821 actgtacccc ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac
     5881 tgctgtctta agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt
     5941 attcatagat tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga
     6001 tcctgctgct tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa
     6061 agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt
     6121 attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag
     6181 tatttgtcta cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag
     6241 ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa
     6301 ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc
     6361 atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc
     6421 ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg
     6481 tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac
     6541 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
     6601 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
     6661 agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc
     6721 ttatatgcag gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg
     6781 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     6841 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     6901 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     6961 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     7021 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     7081 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     7141 gtcaataatc aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     7201 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     7261 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     7321 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     7381 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     7441 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     7501 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     7561 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     7621 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     7681 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7741 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7801 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7861 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7921 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7981 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     8041 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     8101 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     8161 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     8221 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     8281 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     8341 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     8401 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     8461 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8521 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8581 gtgtcttacc gg
//
