LOCUS       sequence-408099-        8409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, LCLAT1_GREB1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8409)
  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 8409)
  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_LCLAT1_GREB10
FEATURES             Location/Qualifiers
     source          1..8409
                     /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(3170..3880,3881..3883,3884..3886))
                     /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(3212..3233)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3561..3580
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3820..3841
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3883..3892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3968..4171)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3997..4017)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4172..4475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4732..4776)
                     /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    4961..5194
                     /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    5554..5671
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6163..6288
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6335..6592
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6594..6973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7004..7108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7109..7177,7178..7969)
                     /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(7327..7346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8140..8409,1..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 ccggttcaca
     1801 atgaacgtgg acttgcgggc tctgcgggtc ccagtgcgac catttgagtc gcgccattga
     1861 caaagacctt aaaaatcacg ggtttcgcct tgcagatttt aagaagatta tcagggactg
     1921 agactggctc tccgctggca acaactggga aggtgaccga tgcaggacct accaggccca
     1981 cctgcggaac acacgacatc ccagcacttt catattttgc acggtttccg cctccctgcg
     2041 ggcagccacc atcgggtgca gagggagatt caggactcca gcctttgtgg cgttttttag
     2101 gaggtcctga gttgctaaga atgcccaacg cggacggacg tggtaaggca agtaagttgt
     2161 tcttggctaa ctgttggcat ttgggagagt ccttcccatt gaatactgct ggacccatag
     2221 cagcgtttaa cgatgggtgg tcactcgcgg gacctgcttg ttgcgcgccc atcagaatgg
     2281 aggggttcgt cccaggaacc ggttcgctag gaaaggccgc tgtgctctcc aaagccggaa
     2341 acagggagga gctgcaggtc gaagccggga tctgacgaga acggaactcc gaccctttcc
     2401 agcagattaa cggccccttt gtcagagtac cctgtgcatt gcgtaccaag tagtacttta
     2461 aatgtttctg tttcttaggc tgtgtggtaa gtttcaagtt aatatgatta ctgaactcgg
     2521 taaaatagca aaaacccttt ttgccacagc cgacgcaatt accggaaaac ccaagaagag
     2581 cgttatgtcc attgtcatca ggcagaaaac gcttgtccac ggcacacact aaaagatgat
     2641 ccggtaagct gggactcttc actccaacca acaaaaaacc agcaggcaca tccatcggct
     2701 cgttcgagat ggataccagg cgcaaatctt tcccggcctg gcagaatcca tcggtcgtac
     2761 aacaaccttc gggcaatggg tgcagttgga acgggttagg tggtccattg gtctcaagcc
     2821 cgccttcgcc ctcctcttct tcttcctcgt tatcgacacg cgagcccccc tcacgtaaac
     2881 gatcaacgac aaatgtgaat cccgtagtac gggggtgaag cacgtattca tatttttgaa
     2941 gtccattttt ttccgcaaaa gcattggagc gcgacttcga attttcggtc agatcggtac
     3001 cttcgggaaa aatcaataat tgcaacggtt cgtgaatgtc acagaagtaa tcgatcatat
     3061 cctcgaaatg actcttgtca tccttccact tgcgatggat aaagatataa gctgcggctt
     3121 gcattccccg ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct
     3181 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     3241 gcttctcgtt ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca
     3301 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     3361 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     3421 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     3481 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     3541 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     3601 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     3661 ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca
     3721 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     3781 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     3841 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg
     3901 gatcccgggc ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca
     3961 ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     4021 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     4081 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     4141 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     4201 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     4261 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     4321 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     4381 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     4441 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     4501 ctaatgaccc cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg
     4561 tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct
     4621 attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat
     4681 tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc
     4741 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
     4801 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
     4861 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
     4921 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
     4981 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
     5041 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
     5101 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
     5161 tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg
     5221 caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt
     5281 atatttatat aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact
     5341 aaacttttcc atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat
     5401 ctgttttcca taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact
     5461 attatcttgt attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca
     5521 aagtggattt ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt
     5581 ttgtaattct ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact
     5641 gtacccccca atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc
     5701 tgtcttaaga tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt
     5761 catagattct actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc
     5821 tgctgcttgc tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc
     5881 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
     5941 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
     6001 ttgtctacag ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc
     6061 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
     6121 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
     6181 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
     6241 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt
     6301 ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa
     6361 aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga
     6421 agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc
     6481 tcccaggctc agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta
     6541 tatgcaggat ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta
     6601 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     6661 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     6721 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     6781 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     6841 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     6901 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     6961 aataatcaat gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7021 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7081 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7141 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7201 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7261 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7321 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7381 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7441 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7501 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7561 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7621 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7681 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7741 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7801 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7861 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7921 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7981 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8041 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     8161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     8221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8281 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     8341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     8401 tcttaccgg
//
