LOCUS       sequence-474364-        8686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of EGFP-IRES-NUP98-KDM5A-KS-FtoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8686)
  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 8686)
  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-IRES-NUP98-KDM5A-KS-FtoA
FEATURES             Location/Qualifiers
     source          1..8686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..583
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(177..194)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     404..423
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             2907..3421
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3583..3599
                     /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(3607..3613,3614..3631,3632..3637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3652..3673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3790..3807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3961..4549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4041..4060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4720..5511,5512..5580))
                     /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     5343..5362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5581..5685)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5753..5771
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5809..5831)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5931..5950
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6144..6166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6294..6313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6513..7029
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7093..7434
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7501..7917
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7847..7869
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7885..7901
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      7938..7947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7944..7946,7947..7949,7950..8663)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7989..8010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8250..8269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8597..8618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
       61 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
      121 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
      181 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
      241 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
      301 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
      361 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
      421 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
      481 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
      541 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttacc ggtgccacca
      601 tgtttaacaa atcatttgga acaccctttg ggggtggcac aggtggcttt ggcacaactt
      661 caacatttgg acagaatact ggctttggca ctactagtgg aggggcattt ggaacatctg
      721 catttggttc tagcaacaat actggaggcc tctttggaaa ttcacagact aaaccaggag
      781 gattgtttgg aaccagttca tttagccagc cagctacctc cacaagcact ggctttgggt
      841 ttggtacgtc aacaggaaca gcaaatacct tgtttggaac tgcaagcaca gggaccagtc
      901 tcttctcatc ccaaaacaat gcctttgcac aaaataaacc aactggcttt ggcaattttg
      961 gaaccagtac tagcagtgga ggactctttg gaaccacaaa taccacctct aatccttttg
     1021 gcagcacatc tggctccctc tttgggccaa gtagttttac agctgctcct actgggacta
     1081 ctattaaatt taaccctcca actggtacag atactatggt caaagctgga gttagcacta
     1141 acataagtac caagcaccag tgtattactg ctatgaaaga atatgaaagc aagtcactag
     1201 aggaacttcg tttagaggat tatcaggcta acaggaaggg cccacagaac caggtgggag
     1261 caggtaccac aactggcttg tttgggtctt ctccagccac ttccagcgca acaggactct
     1321 tcagctcctc caccactaat tcaggctttg catatggtca gaacaaaact gcctttggaa
     1381 ctagtacaac tggatttgga acaaatccag gtggtctctt tggccaacag aatcagcaga
     1441 ctaccagcct cttcagcaaa ccatttggcc aggctacaac cacccagaac actggctttt
     1501 cctttggtaa taccagcacc ataggacagc caagcaccaa caccatggga ttatttggag
     1561 taacccaagc ctcacagcct ggaggtcttt ttgggacagc tacaaacacc agcactggga
     1621 cagcatttgg aacaggaaca ggtctctttg ggcagaccaa tactggattt ggtgctgttg
     1681 gttcgaccct gtttggcaat aacaagctta ctacatttgg aagcagcaca accagtgcac
     1741 cttcatttgg tacaaccagt ggcgggctct ttggttttgg cacaaatacc agtgggaata
     1801 gtatttttgg aagtaaacca gcacctggga ctcttggaac tgggcttggt gcaggatttg
     1861 gaacagctct tggtgctgga caggcatctt tgtttgggaa caaccaacct aagattggag
     1921 ggcctcttgg tacaggagcc tttggggccc ctggatttaa tactacgaca gccactttgg
     1981 gctttggagc cccccaggcc ccagtagctt tgacagatcc aaatgcttct gctgcccagc
     2041 aggctgttct ccagcagcac atcaatagtc taacatactc accttttgga gactctcctc
     2101 tcttccggaa tccgatgtca gaccctaaga agaaggaaga ggatgacagc atggaagaga
     2161 aaccactaaa agtgaaagga aaggactctt cagagaagaa acggaaacgg aagctagaaa
     2221 aggtagagca actttttgga gaaggaaaac agaagtccaa ggagttaaag aaaatggaca
     2281 aacctagaaa gaagaaatta aaattaggtg cagacaaatc aaaggagctg aataaactgg
     2341 ccaagaaact agcaaaagaa gaagagagaa agaaaaagaa ggagaaggct gctgcagcca
     2401 aagttgaact tgtgaaagag agcactgaaa agaaaagaga gaaaaaggtg ctggacatcc
     2461 cctcaaagta tgactggtca ggagcagagg agtctgatga tgagaatgct gtgtgcgcag
     2521 cacagaactg ccaaaggccc tgcaaggaca aggtagactg ggtacaatgt gatggtggct
     2581 gtgatgagtg gtttcatcaa gtttgtgtgg gtgtatctcc agaaatggct gaaaatgaag
     2641 attacatctg tataaactgt gcaaagaagc aggggccagt tagcccaggt ccagcaccac
     2701 ctccttcctt cataatgagc tacaaactac caatggagga tcttaaagag accagtatga
     2761 tgatgatgaa taagttggtt ctagcgcagg ctgctgcttc ctgtctatga aattcgccag
     2821 cacagtgctc gagcacggac atcgacctgc agccaagctt atcgataaaa taaaagattt
     2881 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     2941 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     3001 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     3061 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     3121 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     3181 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     3241 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     3301 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     3361 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     3421 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag
     3481 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt
     3541 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5701 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5761 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5821 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5881 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5941 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6001 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6061 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6121 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
     6181 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     6241 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     6301 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     6361 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
     6421 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     6481 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     6541 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     6601 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     6661 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     6721 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     6781 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     6841 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     6901 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     6961 cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg
     7021 tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc
     7081 ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt
     7141 tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc
     7201 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     7261 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     7321 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     7381 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     7441 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     7501 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     7561 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     7621 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     7681 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     7741 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     7801 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     7861 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
     7921 attgaagatc aacgtacgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     7981 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg
     8041 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     8101 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     8161 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     8221 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     8281 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     8341 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     8401 tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     8461 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     8521 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     8581 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     8641 gcatggacga gctgtacaag taatgaggat ccgcggccgc actaga
//
