LOCUS       sequence-406579-        8220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, ASPSCR1_METRNL from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8220)
  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 8220)
  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_ASPSCR1_METRNL
FEATURES             Location/Qualifiers
     source          1..8220
                     /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(2981..3691,3692..3694,3695..3697))
                     /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(3023..3044)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3372..3391
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3631..3652
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3694..3703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3779..3982)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3808..3828)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3983..4286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4543..4587)
                     /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    4772..5005
                     /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    5365..5482
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5974..6099
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6146..6403
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6405..6784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6815..6919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6920..6988,6989..7780)
                     /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(7138..7157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7951..8220,1..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 ccggttcaat
     1801 cagtcccgac ttcgcatgga ttcaacccgc gttcttgtgc gtcacggtac atacgttgga
     1861 aatctttgaa acggggggca catcccaagc gcgcttctcc aaaatgcatg tgtccggtga
     1921 aaagaaagtc gccgtgtccg ggacggactc cgcactccag aagcgtacgg acacgcccct
     1981 gccaatggcc gtcaccctcc ggaactggtt caaagacgcg gcttttctgg cggtataaac
     2041 gagacacacg caagtggata gcggaatctt ggcgctcagg ttcgtgggtt acttgttgaa
     2101 tcgatccacg gaccgcaaaa tcgctagtac atacagccaa cagtacttcc gtgtctgagc
     2161 aggggcggca tggagcacta agttcatgca gatcggaggc gcggtgacgg cgaaccaatt
     2221 cgtattgaaa tcccgtggtg cggcgtccga tatcctgttg gggggtagct tccacaaaca
     2281 atcctccctg ttctaaaccg aaacactgga cgcgtccagg gcggccgtca ccatctggca
     2341 ccagtaaacg cagttccccg gttttctcca gataaatatt agctccagag gagtcggtaa
     2401 acgagcggat gcacacggtc agatggcgag ccggactaaa ggtattcggg cgaagattaa
     2461 caatcagggc gcctgtaggg tacatccatt ctactgcgcc cgccgcacag cgcagataca
     2521 cctgctcaac ttctttgcgg tgtgcttcgt gggtcaaacc gctaatctgc ggaaagtgtg
     2581 ataacaactc ccaaagcgtt tggccagaac agaatgagtc ctgcaaacgt gaaccatcgt
     2641 ccaactgaag cgcaatacgc accatgttct ctggaccctc acgactacgt gaagctggga
     2701 ccatttcaag tttagcgttg ttaggaaggt tcgcgaaacg ccactgaagt gacaggtcta
     2761 acacagagcg ttgaaacttc aaatcgtatt cgcatgggtt gaaatcttga cggcggcaag
     2821 tatcttccaa aacctgcaac agaactgttg aaggcgtgac cttgacggta tgacgacggc
     2881 catttggtgc caatacagag acagcagacc cacctcctcc cgcaggagct gccattcccc
     2941 gggaagcggc cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc
     3001 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     3061 tggggtcttt gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     3121 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     3181 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     3241 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     3301 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     3361 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     3421 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     3481 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     3541 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     3601 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     3661 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg
     3721 cccgcggtac cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag
     3781 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     3841 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     3901 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     3961 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     4021 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4081 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4141 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4201 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4261 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4321 ccgtaattga ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg
     4381 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
     4441 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
     4501 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
     4561 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
     4621 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
     4681 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
     4741 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
     4801 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
     4861 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
     4921 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
     4981 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
     5041 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
     5101 taattcactt ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc
     5161 catgttctaa tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc
     5221 ataatcccta atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg
     5281 tattactact gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt
     5341 tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc
     5401 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc
     5461 aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag
     5521 atgttcagcc tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc
     5581 tactactcct tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg
     5641 ctgtgctttt ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt
     5701 gtcttttatc tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga
     5761 tctaagttct tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca
     5821 gccttctgat gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc
     5881 ccatactata tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc
     5941 ccatcgcgat ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt
     6001 ctagcctccg ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc
     6061 cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc
     6121 tttcaagtcc ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg
     6181 agggatctct agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa
     6241 ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct
     6301 cagatctggt ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga
     6361 tctgagggct cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg
     6421 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     6481 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     6541 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     6601 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     6661 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     6721 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     6781 tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6901 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6961 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7021 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7081 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7141 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7201 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7261 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7321 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7381 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7441 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7501 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7561 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7621 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7681 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7741 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7801 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7861 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7921 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7981 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8041 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     8101 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8161 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
//
