LOCUS       sequence-407492-        8754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, TTC6_MIPOL1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8754)
  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 8754)
  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_TTC6_MIPOL1
FEATURES             Location/Qualifiers
     source          1..8754
                     /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(3515..4225,4226..4228,4229..4231))
                     /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(3557..3578)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3906..3925
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4165..4186
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4228..4237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4313..4516)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4342..4362)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4517..4820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5077..5121)
                     /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    5306..5539
                     /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    5899..6016
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6508..6633
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6680..6937
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6939..7318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7349..7453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7454..7522,7523..8314)
                     /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(7672..7691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8485..8754,1..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 tcacagtgcg catagtgcct gtaccaactt tcttgcgcaa gacgtccaca aggcgctcta
     1861 acttttggac tttctcattc gccacttggc tggcttcacg agcgtgttga agttgtaatt
     1921 ccatattagc acgttcggtc agagcctgct gtaattgggt tgcaagctcc tcgttttgtt
     1981 gctgcgtaat tgacacgatg ttctcgcgat ttttcagtgc ttcctcgtaa gtcgacagag
     2041 tgtaattgaa ttgatccttc agattctctt cttggctaag ggatttatgt aagtttttat
     2101 gcagcttttt gtcacaaagc ggttttgatg acttgcgcgg acgcaaactc ttgcgcttgc
     2161 gcatgcgctt gatctccttc ccctcaatct gcagaaactc aggtgatttc gccgtcagcg
     2221 atacccattt cgcctccttg gagtcactag ttggctgtcc gtcctctaag ggctttgaaa
     2281 gttcctctgc tggttggacc tgggagattt cctgtggctt gaacacgata gactcggctt
     2341 cactcggtcc ccactcgatg tcctcctgtt cgatctgcag gatggaggac attgtctctg
     2401 acaattcttt atgaacactc atttgtgact ccttggcccc catttgaaac ttgtattcgg
     2461 cttggatttc tggcgtctcc gcttgcatag gggacgtttt taaatacatt ttacccatta
     2521 agctgatatc ctccattgtg atatcgtaat tctggcccag gacactgcgg atcagttctt
     2581 tgatggtctg gtcagatgct aattgtgctt cggactgaag actcgcgatg atttcctcga
     2641 tagaggtggg aatcacacga gaaggaagca gtgcctgcca agcagcttcg cgcgcatcag
     2701 atggtggcca tgattcctgc tccccctgtg cgcctaaacc agcagcctcg cgtgcctcac
     2761 gggcccccga ctcagacaca aaattactgc gaatacggac tttacgacgc ccgctggaag
     2821 cttccatata accgtcttcg gcggaaacac ttgatgcctt acgttctttg gccatacacg
     2881 ggccagccag aggagctacc tcttgagact gttcacgttc acgacctaaa tggaatacgc
     2941 ggcgtggtgc agcgtggcga gatgactcca gcacaccacg cgcgcgacgc ttaaccggtg
     3001 gctctggtgg cggtaaaagg acgaccgggc gggcagtacc aaagccactt gcgattacgg
     3061 gaggtttttt taaggcttgg tgacgtaaga aggctgatga gcgcaaataa aaatcacgag
     3121 gacgaaaaga gcgcgtctta ccaggtgcag cggcttcccc taacccgctc ggacgcggag
     3181 cacctccaag cacttcttgc agaagtgcag cggcgctcat tgccggacct tttaacgatg
     3241 ggtaacggcg cgatgtacgg gctgccgcgc atgatacgcg ggccgggcgt gcaggacctg
     3301 gggcatgtaa ggaatgcact ggcgactcat caactgctgg cttcgaggcc aacttctgct
     3361 taaagcgaag gaagtctttt ttagtctcct ggcgtacctt ctccagttct ttgaacatat
     3421 acgactcttc cttatatttc agtccgaagt ggcgcggaat cgtactcatt ccccgggaag
     3481 cggccgcact tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag
     3541 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     3601 ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     3661 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     3721 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     3781 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     3841 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     3901 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     3961 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     4021 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     4081 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     4141 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     4201 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg
     4261 gtaccgtcga ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc
     4321 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     4381 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     4441 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     4501 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     4561 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     4621 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     4681 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     4741 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     4801 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     4861 ttgattacta ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg
     4921 gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag
     4981 tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga ataattttat
     5041 ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc
     5101 caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat
     5161 taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat
     5221 ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag
     5281 ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc
     5341 caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg
     5401 cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct
     5461 ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa
     5521 cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt
     5581 tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt tatataattc
     5641 acttctccaa ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt
     5701 ctaatcctca tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc
     5761 cctaatgatc tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac
     5821 tactgcccct tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct
     5881 gtccctgtaa taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt
     5941 ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc
     6001 cccttttctt ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc
     6061 agcctgatct cttacctgtc ctataatttt ctttaattct ttattcatag attctactac
     6121 tccttgactt tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc
     6181 ttttttctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt
     6241 tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag
     6301 ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc
     6361 tgatgtttct aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac
     6421 tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg
     6481 cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc
     6541 tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg
     6601 cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa
     6661 gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat
     6721 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
     6781 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
     6841 tggtctaacc agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag
     6901 ggctcgccac tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat
     6961 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     7021 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     7081 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     7141 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     7201 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     7261 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga
     7321 cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7381 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7441 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7501 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7561 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7621 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7681 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7741 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7801 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7861 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7921 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7981 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8041 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8101 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8161 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8221 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8281 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8341 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8401 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8461 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8521 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8581 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8641 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8701 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgg
//
