LOCUS       sequence-408144-        7779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MLLT10_PICALM from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7779)
  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 7779)
  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_MLLT10_PICALM
FEATURES             Location/Qualifiers
     source          1..7779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     421..438
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    506..640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(530..549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     584..603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(646..841)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      660..795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    900..921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(936..941,942..959,960..966)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    974..990
                     /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     979..1001
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     998..1014
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     998..1014
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1357..1376
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1375..1430
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1476..1495
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(2488..3198,3199..3201,3202..3204))
                     /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(2530..2551)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2879..2898
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3138..3159
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3201..3210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3286..3489)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3315..3335)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3490..3793
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4050..4094)
                     /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    4279..4512
                     /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    4872..4989
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5481..5606
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5653..5910
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5912..6291
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6322..6426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6427..6495,6496..7287)
                     /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(6645..6664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7458..7779,1..267)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
       61 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      301 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      361 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      421 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      481 cgttggccga ttcattaatg cagcggatcc agacatgata agatacattg atgagtttgg
      541 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      601 tgctttattt gtaaccatta taagctgcaa taaacaagtt agctttttgc aaaagcctag
      661 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
      721 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
      781 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
      841 cggatccgct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      901 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
      961 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     1021 tacgccaagc ttgggctgca ggtcgaggga tctccataag agaagaggga cagctatgac
     1081 tgggagtagt caggagagga ggaaaaatct ggctagtaaa acatgtaagg aaaattttag
     1141 ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa aaaatctaac ctcaagtcaa
     1201 ggcttttcta tggaataagg aatggacagc agggggctgt ttcatatact gatgacctct
     1261 ttatagccaa cctttgttca tggcagccag catatgggca tatgttgcca aactctaaac
     1321 caaatactca ttctgatgtt ttaaatgatt tgccctccca tatgtccttc cgagtgagag
     1381 acacaaaaaa ttccaacaca ctattgcaat gaaaataaat ttcctttatt agccagaagt
     1441 cagatgctca aggggcttca tgatgtcccc ataatttttg gcagagggaa aaagatctgc
     1501 tagctcgagc agctgaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
     1561 agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagatct
     1621 tgtcttcttt gggagtgaat tagcccttcc agtcccccct tttcttttaa aaagtggcta
     1681 agatctacag ctgccttgta agtcattggt cttaaaggta ccgtaatctt atctagacta
     1741 catgaactga atcttctttt ctttttcgtg gtgtttgtcc tggctgtgcg aggacgagtc
     1801 tgacagcgac tggtcatagg agcgattgga cccgcgtttt gactttttca gtttggaaaa
     1861 gtggtactta cagtagccgc agtactgtac gttatccgct ccattccctt cttcctcaca
     1921 taacaagccc gcaaactgtg cgcatgtaac gtgaaaagcc tggcggcagc catgcttatt
     1981 acaggtcatg caagcgcccg tagcggcctt actctcacgc ccttgctcgt cacagatata
     2041 acacgtcttg ttgtagcgat cgtgtggcac actttgaagt acaatgggtt ccatagtgct
     2101 gacgttggcg aactgaactt ccgggatata cagtgcacaa acaacatgcg cccaccctcc
     2161 gttatcggta cgtttaagag cgccgtcctt atgggggcac aactcgcaac gtacgcgtgc
     2221 tgcgcgctct tgagattcac acttgcggca aaaccagggg cctgtcggaa cttggacgat
     2281 gccgtaacag gcttggtgga cggccactga acaaccgtgc ccgtcgcaat agaccaaggg
     2341 attttccgcc cacccacgct catcgctaca gacgcagcag cctccaatca tctctttcat
     2401 ggaatgggaa acctcatctt caagtgatac gggacgatcg ctagagacca tagcggccgc
     2461 acttcccccg gagcctccgc ttccgccctt gtacagctcg tccatgccga gagtgatccc
     2521 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     2581 cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     2641 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     2701 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
     2761 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     2821 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     2881 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     2941 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     3001 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     3061 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     3121 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     3181 cagctcctcg cccttgctca ccatggtggc gaccggtgga tcccgggccc gcggtaccgt
     3241 cgactgcaga attcggttca ctaaacgagc ccggttcact aaacgagctc tgcttatata
     3301 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     3361 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     3421 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     3481 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3541 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3601 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3661 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3721 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3781 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3841 ctattaataa ctagcacgcg ttcgaagctt gagctcggtc ccctcggggt tgggaggtgg
     3901 gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga aagtacagca
     3961 aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt tatataccac
     4021 agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa ttccaataat
     4081 tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg tattaagctt
     4141 gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca aatctgttcc
     4201 agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat gagttttcca
     4261 gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac agccaggatt
     4321 cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg ttgcgcctca
     4381 atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg tctggcctgt
     4441 accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg
     4501 aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct ctttgccttg
     4561 gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa ttcacttctc
     4621 caattgtccc tcatatctcc tccttatggt gttttactaa acttttccat gttctaatcc
     4681 tcatcctgtc tacttgccac acaatcatca cctgccatct gttttccata atccctaatg
     4741 atctttgctt ttcttcttgg cactactttt atgtcactat tatcttgtat tactactgcc
     4801 ccttcacctt tccagaggag ctttgctggt cctttccaaa gtggatttct gctgtccctg
     4861 taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
     4921 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     4981 cttttaaaat tgtggatgaa tactgccatt tgtactgctg tcttaagatg ttcagcctga
     5041 tctcttacct gtcctataat tttctttaat tctttattca tagattctac tactccttga
     5101 ctttggggat tgtagggaat ttccaggtct gaagatcctg ctgcttgctg tgcttttttc
     5161 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
     5221 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
     5281 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
     5341 tctaacaggc cagtattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
     5401 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
     5461 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
     5521 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
     5581 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
     5641 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     5701 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     5761 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     5821 accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct gagggctcgc
     5881 cactccccag tcccgcccag gccacgcctc ccatggtaat agcgatgact aatacgtaga
     5941 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     6001 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     6061 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     6121 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     6181 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     6241 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt cgacccaggt
     6301 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6361 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6421 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6481 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6541 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6601 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6661 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6721 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6781 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6841 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6901 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6961 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7021 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7081 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7141 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7201 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7261 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7321 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7381 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7441 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7501 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7561 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7621 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7681 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7741 cgatagttac cggataaggc gcagcggtcg ggctgaacg
//
