LOCUS       sequence-474377-        8190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of EGFP-NUP98-KDM5A-KS-FtoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8190)
  AUTHORS   Zhang Y, Stoppelkamp I, Fernandez-Pernas P, Allram M, Charman M, 
            Magalhaes AP, Piedavent-Salomon M, Sommer G, Sung YC, Meyer K, Grams
            N, Halko E, Dongre S, Meierhofer D, Malszycki M, Ilik IA, Aktas T, 
            Kraushar ML, Vastenhouw N, Weitzman MD, Grebien F, Niskanen H, Hnisz
            D
  TITLE     Probing condensate microenvironments with a micropeptide killswitch.
  JOURNAL   Nature. 2025 Jun 4. doi: 10.1038/s41586-025-09141-5.
  PUBMED    40468084
REFERENCE   2  (bases 1 to 8190)
  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: "Nature. 
            2025 Jun 4. doi: 10.1038/s41586-025-09141-5."
COMMENT     Sequence Label: mscv_EGFP-NUP98-KDM5A-KS-FtoA
FEATURES             Location/Qualifiers
     source          1..8190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     471..490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(709..813)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     881..899
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(937..959)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1059..1078
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1272..1294
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1422..1441
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             1641..2157
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2221..2562
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2629..3045
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2975..2997
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3013..3029
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      3066..3075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3072..3074,3075..3077,3078..3788)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3072..3074,3075..3077,3078..3788)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3117..3138)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3378..3397)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3725..3746
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3789..3830
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3834..3875
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3879..3920
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     LTR             6225..6739
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6901..6917
                     /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(6925..6931,6932..6949,6950..6955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6970..6991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7108..7125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7279..7867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7359..7378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8038..8190,1..639,640..708))
                     /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"
ORIGIN
        1 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
       61 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
      421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
      481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
      541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
      601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
      661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
      721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
      841 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      901 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      961 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     1021 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     1081 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     1141 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1201 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1261 gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag
     1321 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     1381 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     1441 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     1501 cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa
     1561 caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt
     1621 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     1681 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
     1741 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     1801 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     1861 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     1921 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     1981 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     2041 ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct
     2101 gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg
     2161 aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt
     2221 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc
     2281 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt
     2341 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
     2401 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
     2461 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     2521 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
     2581 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
     2641 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     2701 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     2761 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     2821 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     2881 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     2941 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     3001 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tgaagatcaa
     3061 cgtacgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3121 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     3181 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3241 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3301 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3361 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3421 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3481 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3541 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3601 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3661 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3721 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3781 tgtacaaggg aaagccaatc ccaaaccctc tgctgggact ggacagcacc tccggcaagc
     3841 ctatccctaa ccccctgctc gggctggaca gtacgtccgg taagcccatc cccaacccac
     3901 tcctgggact cgatagtaca tttaacaaat catttggaac accctttggg ggtggcacag
     3961 gtggctttgg cacaacttca acatttggac agaatactgg ctttggcact actagtggag
     4021 gggcatttgg aacatctgca tttggttcta gcaacaatac tggaggcctc tttggaaatt
     4081 cacagactaa accaggagga ttgtttggaa ccagttcatt tagccagcca gctacctcca
     4141 caagcactgg ctttgggttt ggtacgtcaa caggaacagc aaataccttg tttggaactg
     4201 caagcacagg gaccagtctc ttctcatccc aaaacaatgc ctttgcacaa aataaaccaa
     4261 ctggctttgg caattttgga accagtacta gcagtggagg actctttgga accacaaata
     4321 ccacctctaa tccttttggc agcacatctg gctccctctt tgggccaagt agttttacag
     4381 ctgctcctac tgggactact attaaattta accctccaac tggtacagat actatggtca
     4441 aagctggagt tagcactaac ataagtacca agcaccagtg tattactgct atgaaagaat
     4501 atgaaagcaa gtcactagag gaacttcgtt tagaggatta tcaggctaac aggaagggcc
     4561 cacagaacca ggtgggagca ggtaccacaa ctggcttgtt tgggtcttct ccagccactt
     4621 ccagcgcaac aggactcttc agctcctcca ccactaattc aggctttgca tatggtcaga
     4681 acaaaactgc ctttggaact agtacaactg gatttggaac aaatccaggt ggtctctttg
     4741 gccaacagaa tcagcagact accagcctct tcagcaaacc atttggccag gctacaacca
     4801 cccagaacac tggcttttcc tttggtaata ccagcaccat aggacagcca agcaccaaca
     4861 ccatgggatt atttggagta acccaagcct cacagcctgg aggtcttttt gggacagcta
     4921 caaacaccag cactgggaca gcatttggaa caggaacagg tctctttggg cagaccaata
     4981 ctggatttgg tgctgttggt tcgaccctgt ttggcaataa caagcttact acatttggaa
     5041 gcagcacaac cagtgcacct tcatttggta caaccagtgg cgggctcttt ggttttggca
     5101 caaataccag tgggaatagt atttttggaa gtaaaccagc acctgggact cttggaactg
     5161 ggcttggtgc aggatttgga acagctcttg gtgctggaca ggcatctttg tttgggaaca
     5221 accaacctaa gattggaggg cctcttggta caggagcctt tggggcccct ggatttaata
     5281 ctacgacagc cactttgggc tttggagccc cccaggcccc agtagctttg acagatccaa
     5341 atgcttctgc tgcccagcag gctgttctcc agcagcacat caatagtcta acatactcac
     5401 cttttggaga ctctcctctc ttccggaatc cgatgtcaga ccctaagaag aaggaagagg
     5461 atgacagcat ggaagagaaa ccactaaaag tgaaaggaaa ggactcttca gagaagaaac
     5521 ggaaacggaa gctagaaaag gtagagcaac tttttggaga aggaaaacag aagtccaagg
     5581 agttaaagaa aatggacaaa cctagaaaga agaaattaaa attaggtgca gacaaatcaa
     5641 aggagctgaa taaactggcc aagaaactag caaaagaaga agagagaaag aaaaagaagg
     5701 agaaggctgc tgcagccaaa gttgaacttg tgaaagagag cactgaaaag aaaagagaga
     5761 aaaaggtgct ggacatcccc tcaaagtatg actggtcagg agcagaggag tctgatgatg
     5821 agaatgctgt gtgcgcagca cagaactgcc aaaggccctg caaggacaag gtagactggg
     5881 tacaatgtga tggtggctgt gatgagtggt ttcatcaagt ttgtgtgggt gtatctccag
     5941 aaatggctga aaatgaagat tacatctgta taaactgtgc aaagaagcag gggccagtta
     6001 gcccaggtcc agcaccacct ccttccttca taatgagcta caaactacca atggaggatc
     6061 ttaaagagac cagtatgatg atgatgaata agttggttct agcgcaggct gctgcttcct
     6121 gtctatgaaa ttcgccagca cagtgctcga gcacggacat cgacctgcag ccaagcttat
     6181 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     6241 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     6301 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     6361 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     6421 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     6481 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     6541 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     6601 agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     6661 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     6721 gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg
     6781 taggagtcga atattaagta atcctgactc aattagccac tgttttgaat ccacatactc
     6841 caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa
     6901 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6961 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7021 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7081 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7681 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8101 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8161 tgctgcaatg ataccgcgag acccacgctc
//
