LOCUS       sequence-407491-        8028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, WASL_VPS13D from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  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 8028)
  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_WASL_VPS13D
FEATURES             Location/Qualifiers
     source          1..8028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(123..227)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        258..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             639..896
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    943..1068
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1560..1677
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2037..2270
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2455..2499
                     /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"
     enhancer        2756..3059
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3060..3263
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3214..3234
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3339..3348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3345..3347,3348..3350,3351..4061)
                     /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(3390..3411)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3651..3670)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3998..4019
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5303..5322)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5368..5423
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5422..5441)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5784..5800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5784..5800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5797..5819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5808..5824
                     /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)."
     promoter        complement(join(5832..5838,5839..5856,5857..5862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5877..5898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5957..6152
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6003..6138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6158..6292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6195..6214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6249..6268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6360..6377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6531..7119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6611..6630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7290..8028,1..53,54..122))
                     /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     7913..7932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
       61 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
      121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
      181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
      241 aaagtgccac ctgggtcgac attgattatt gactagttat taatagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatggga ggcgtggcct gggcgggact
      661 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
      721 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      781 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      841 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      901 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      961 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1021 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1081 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1141 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatactggc
     1201 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1261 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1321 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1381 aaaagtaaga aaaaagcaca gcaagcagca ggatcttcag acctggaaat tccctacaat
     1441 ccccaaagtc aaggagtagt agaatctatg aataaagaat taaagaaaat tataggacag
     1501 gtaagagatc aggctgaaca tcttaagaca gcagtacaaa tggcagtatt catccacaat
     1561 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     1621 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     1681 gtttattaca gggacagcag aaatccactt tggaaaggac cagcaaagct cctctggaaa
     1741 ggtgaagggg cagtagtaat acaagataat agtgacataa aagtagtgcc aagaagaaaa
     1801 gcaaagatca ttagggatta tggaaaacag atggcaggtg atgattgtgt ggcaagtaga
     1861 caggatgagg attagaacat ggaaaagttt agtaaaacac cataaggagg agatatgagg
     1921 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1981 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     2041 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     2101 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     2161 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     2221 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     2281 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     2341 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     2401 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     2461 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     2521 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2581 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2641 tttcagaccc acctcccaac cccgagggga ccgagctcaa gcttcgaacg cgtgctagtt
     2701 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2761 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2821 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2881 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2941 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3001 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     3061 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     3121 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3181 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     3241 gggaggtcta tataagcaga gctcgtttag tgaaccgggc tcgtttagtg aaccgaattc
     3301 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg
     3361 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3421 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3481 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     3541 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3601 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3661 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3721 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3781 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3841 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3901 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3961 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4021 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggaagc ggaggctccg
     4081 ggggaagtgc ggccgcttcc cggatgtcgt ccgtccagca acagccgccc cctcctcgcc
     4141 gtgttactaa tgtaggaagc cttttactta ccccgcagga gaatgaaagt ctttttacat
     4201 ttcttggtaa aaaatgtgtt tttgctggaa ccttgtcgga tgggcttggg aagactatgg
     4261 acaaccgtca tcagtccgag cgtgaataca ttcgctatca cgcggctacc tcaggagaac
     4321 atcttgtggc tggtatccac gggttggcac acgggattat cggcgggctt acctcggtta
     4381 ttacgagcac cgtggaagga gtcaaaacag agggtggcgt cagcggcttt atttcaggtc
     4441 tgggaaaagg tctggtcggg acggtcacca aacccgtggc cggggcgttg gacttcgcta
     4501 gcgaaactgc acaagcggtc cgtgacacag ctactctttc tggcccacgc actcaagcac
     4561 agcgcgtgcg taagccgcgc tgttgtaccg gtccgcaggg attactgccc cgctacagcg
     4621 agagccaagc agaaggtcaa gaacagttat tcaaacttac agacaatatc caagacgagt
     4681 tcttcatcgc agtcgagaac attgattcgt actgtgtgct tatcagcagt aaggcagttt
     4741 attttcttaa atcaggtgat tacgtggacc gcgaagcaat ctttctggaa gtgaagtacg
     4801 atgaccttta ccactgtctt gtgagcaagg atcatggcaa agtctacgta caagtgacga
     4861 agaaggcggt cagcacttca tcgggagtat ccatcccagg cccttcacat cagaagccca
     4921 tggtgcatgt caagtctgaa gtgctggctg tgaagctgag tcaagagatc aactacgcga
     4981 aaagtttgta ttacgagcaa caactgatgc tgcgtttgtc ggagaatcgc gagcaattag
     5041 agttagatag ttgaaccggt tctagataag attacggtac ctttaagacc aatgacttac
     5101 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     5161 cactcccaaa gaagacaaga tctgcttttt gcctgtactg ggtctctctg gttagaccag
     5221 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     5281 ttcagctgct cgagctagca gatctttttc cctctgccaa aaattatggg gacatcatga
     5341 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     5401 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     5461 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     5521 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     5581 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     5641 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     5701 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     5761 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5821 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5881 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5941 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6001 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6061 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6121 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa
     6181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat
     6301 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6841 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6961 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7201 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7261 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccact
//
