LOCUS       sequence-471769-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of 52K-GFP-KS-FtoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  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 6070)
  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: p52K-GFP-KS-FtoA
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(273..377)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      404..859
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(491..510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     701..722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    866..987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(877..896)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     931..950
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1170..1880,1881..1883))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITLGMDELYK"
     primer_bind     1561..1580
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1820..1841
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3277..3480)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3306..3326)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3481..3784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3959..4547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4039..4058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4876..4923
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4948..4967
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5155..5949)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5155..5949)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5266..5285)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5876..5895
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5984..6070,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5998..6070,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
      301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
      361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
      421 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      481 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      541 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      601 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      661 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      721 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      781 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      841 atattaacgc ttacaattta cgccttaaga tacattgatg agtttggaca aaccacaact
      901 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      961 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1021 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1081 gctgattatg atctagagtc gcggccgctt tacttctata gacaggaggc ggcggcctgc
     1141 gctagaacca acttattcat catcatcatc ttgtacagct cgtccatgcc gagagtgatc
     1201 ccggcggcgg tcacgaactc cttcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     1261 ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     1321 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     1381 cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
     1441 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     1501 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     1561 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     1621 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
     1681 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     1741 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     1801 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     1861 aacagctcct cgcccttgct cactatggtg acgaccggtg gatcccgggc ccgcggtacc
     1921 gtcgactgca gaattcggta ctcgccgtcc tctggctcgt actcatcgtc ctcgtcatat
     1981 tcctccacgc cgccgacgtt gccagcgcgc gcgggtgcca ccgccagccc aggtccggcc
     2041 ccagctgcct ccagggcgcg tcggcttggg gcccagcgca ggtcagcgcc cgcgtcaaag
     2101 taggactcgg cctctctatc gccgctgccc gtgccagcca gggccctttg caggctgtgc
     2161 atcagctcgc ggtcgctgag ctcgcgccgc cggctcacgc tcacggcctt gtggatgcgc
     2221 tcgttgcgat aaacgcccag gtcgtcgctc aaggtaagca ccttcagcgc catgcgcatg
     2281 tagaacccct cgatctttac ctccttgtct atgggaacgt aaggggtatg gtatatcttg
     2341 cgggcgtaaa acttgcccag gctaagcatg gaatagttga tggcggccac cttgtcagcc
     2401 aggctcaagc tgcgctcctg caccactatg ctctgcagga tgtttatcaa atcgagcagc
     2461 cagcggccct cgggctctac tatgtttagc agcgcatccc tgaatgcctc gttgtccctg
     2521 ctgtgctgca ctataaggaa cagctgcgcc atgagcggct tgctatttgg gttttgctcc
     2581 agcgcgctta caaagtccca cagatgcatc agtcctatag ccacctcctc gcgcgccaca
     2641 agcgtacgca cgtggttgtt aaagcttttt tgaaagttaa tctcctggtt caccgtctgc
     2701 tcgtatgcgg ttaccaggtc ggcggccgcc acgtgtgcgc gcgcgggact aatcccggtt
     2761 cgcgcgtcgg gctcaaagtc ctcctcgcgc agcaaccgct cgcgattcag gccatgccgc
     2821 agctcgcgcc ctgcgtggaa ctttcgatcc cgcatctcct cgggctcctc tccctcgcgg
     2881 tcgcgaaaca ggttctgccg cggcacgtac gcctcacgcg tatcacgctt cagctgcacc
     2941 cttgggtgcc gctcaggaga gggcgctcct agccgcgcca ggccctcgcc ctcctccaag
     3001 tccaggtagt gccgggcccg gcgccgcggg ggttcgtaat caccatctgc tgccgcgtca
     3061 accgcggatg tcgcccctcc tgacgcggta ggaggagggg agggtgccct gcatgtctgc
     3121 cgctgctctt gctcttgccg ctgctgagga ggggggcgca tctgccgcag caccggatgc
     3181 atctcgagat ctgagtccgg tagcgctagc ggatctgacg gttcacacta gcggatctga
     3241 cggttcacgc tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca
     3301 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     3361 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     3421 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     3481 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     3541 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     3601 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     3661 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3721 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3781 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3841 actaatgcat ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4381 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4681 caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg
     4741 ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc
     4801 gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     4861 gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg
     4921 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     4981 tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     5041 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     5101 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     5161 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     5221 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     5281 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     5341 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     5401 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     5461 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     5521 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     5581 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     5641 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     5701 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     5761 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     5821 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     5881 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     5941 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc
     6001 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     6061 ctgcataaat
//
