LOCUS       sequence-474432-        8635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of EGFP-IRES-NUP98-KDM5A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8635)
  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 8635)
  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
FEATURES             Location/Qualifiers
     source          1..8635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..582
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(176..193)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     403..422
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             2855..3369
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3531..3547
                     /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(3555..3561,3562..3579,3580..3585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3600..3621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3738..3755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4668..5459,5460..5528))
                     /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     5291..5310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5529..5633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5701..5719
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5757..5779)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5879..5898
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6092..6114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6242..6261
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6461..6977
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7041..7382
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7449..7865
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7795..7817
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7833..7849
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      7886..7895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7892..7894,7895..7897,7898..8611)
                     /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(7937..7958)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8198..8217)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8545..8566
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 caattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
       61 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
      121 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
      181 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
      241 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
      301 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
      361 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
      421 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
      481 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
      541 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttaccg gtgccaccat
      601 gtttaacaaa tcatttggaa caccctttgg gggtggcaca ggtggctttg gcacaacttc
      661 aacatttgga cagaatactg gctttggcac tactagtgga ggggcatttg gaacatctgc
      721 atttggttct agcaacaata ctggaggcct ctttggaaat tcacagacta aaccaggagg
      781 attgtttgga accagttcat ttagccagcc agctacctcc acaagcactg gctttgggtt
      841 tggtacgtca acaggaacag caaatacctt gtttggaact gcaagcacag ggaccagtct
      901 cttctcatcc caaaacaatg cctttgcaca aaataaacca actggctttg gcaattttgg
      961 aaccagtact agcagtggag gactctttgg aaccacaaat accacctcta atccttttgg
     1021 cagcacatct ggctccctct ttgggccaag tagttttaca gctgctccta ctgggactac
     1081 tattaaattt aaccctccaa ctggtacaga tactatggtc aaagctggag ttagcactaa
     1141 cataagtacc aagcaccagt gtattactgc tatgaaagaa tatgaaagca agtcactaga
     1201 ggaacttcgt ttagaggatt atcaggctaa caggaagggc ccacagaacc aggtgggagc
     1261 aggtaccaca actggcttgt ttgggtcttc tccagccact tccagcgcaa caggactctt
     1321 cagctcctcc accactaatt caggctttgc atatggtcag aacaaaactg cctttggaac
     1381 tagtacaact ggatttggaa caaatccagg tggtctcttt ggccaacaga atcagcagac
     1441 taccagcctc ttcagcaaac catttggcca ggctacaacc acccagaaca ctggcttttc
     1501 ctttggtaat accagcacca taggacagcc aagcaccaac accatgggat tatttggagt
     1561 aacccaagcc tcacagcctg gaggtctttt tgggacagct acaaacacca gcactgggac
     1621 agcatttgga acaggaacag gtctctttgg gcagaccaat actggatttg gtgctgttgg
     1681 ttcgaccctg tttggcaata acaagcttac tacatttgga agcagcacaa ccagtgcacc
     1741 ttcatttggt acaaccagtg gcgggctctt tggttttggc acaaatacca gtgggaatag
     1801 tatttttgga agtaaaccag cacctgggac tcttggaact gggcttggtg caggatttgg
     1861 aacagctctt ggtgctggac aggcatcttt gtttgggaac aaccaaccta agattggagg
     1921 gcctcttggt acaggagcct ttggggcccc tggatttaat actacgacag ccactttggg
     1981 ctttggagcc ccccaggccc cagtagcttt gacagatcca aatgcttctg ctgcccagca
     2041 ggctgttctc cagcagcaca tcaatagtct aacatactca ccttttggag actctcctct
     2101 cttccggaat ccgatgtcag accctaagaa gaaggaagag gatgacagca tggaagagaa
     2161 accactaaaa gtgaaaggaa aggactcttc agagaagaaa cggaaacgga agctagaaaa
     2221 ggtagagcaa ctttttggag aaggaaaaca gaagtccaag gagttaaaga aaatggacaa
     2281 acctagaaag aagaaattaa aattaggtgc agacaaatca aaggagctga ataaactggc
     2341 caagaaacta gcaaaagaag aagagagaaa gaaaaagaag gagaaggctg ctgcagccaa
     2401 agttgaactt gtgaaagaga gcactgaaaa gaaaagagag aaaaaggtgc tggacatccc
     2461 ctcaaagtat gactggtcag gagcagagga gtctgatgat gagaatgctg tgtgcgcagc
     2521 acagaactgc caaaggccct gcaaggacaa ggtagactgg gtacaatgtg atggtggctg
     2581 tgatgagtgg tttcatcaag tttgtgtggg tgtatctcca gaaatggctg aaaatgaaga
     2641 ttacatctgt ataaactgtg caaagaagca ggggccagtt agcccaggtc cagcaccacc
     2701 tccttccttc ataatgagct acaaactacc aatggaggat cttaaagaga ccagttagaa
     2761 ttcgccagca cagtgctcga gcacggacat cgacctgcag ccaagcttat cgataaaata
     2821 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     2881 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     2941 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     3001 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     3061 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3121 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3181 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3241 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     3301 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     3361 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga
     3421 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct
     3481 gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg
     3541 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3601 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3661 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3721 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3781 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3841 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3901 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4561 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4621 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4801 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5101 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5401 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5521 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5581 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5641 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5701 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     5761 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     5821 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     5881 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     5941 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     6001 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6061 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
     6121 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6181 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6241 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6301 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
     6361 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     6421 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     6481 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     6541 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     6601 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     6661 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     6721 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     6781 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     6841 cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc
     6901 atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac
     6961 ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac
     7021 cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg
     7081 tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc
     7141 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
     7201 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
     7261 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
     7321 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
     7381 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
     7441 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
     7501 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
     7561 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
     7621 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
     7681 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
     7741 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
     7801 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
     7861 gcgccggaat tgaagatcaa cgtacgccac catggtgagc aagggcgagg agctgttcac
     7921 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     7981 gtctggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     8041 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     8101 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     8161 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     8221 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     8281 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     8341 cgtctatatc atggccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca
     8401 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     8461 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     8521 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     8581 cactctcggc atggacgagc tgtacaagta atgaggatcc gcggccgcac tagag
//
