LOCUS       sequence-471758-        6100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of 52K-GFP-linker-KS-FtoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6100)
  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 6100)
  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-linker-KS-FtoA
FEATURES             Location/Qualifiers
     source          1..6100
                     /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(1200..1910,1911..1913))
                     /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     1591..1610
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1850..1871
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3307..3510)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3336..3356)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3511..3814
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3989..4577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4069..4088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4906..4953
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4978..4997
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5185..5979)
                     /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(5185..5979)
                     /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(5296..5315)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5906..5925
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6014..6100,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6028..6100,1..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 catcatcata gatcctccgc ctccgcttcc tccgccgccc
     1201 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cttcaggacc
     1261 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     1321 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1381 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     1441 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1501 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1561 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     1621 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     1681 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     1741 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     1801 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     1861 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct cactatggtg
     1921 acgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggta ctcgccgtcc
     1981 tctggctcgt actcatcgtc ctcgtcatat tcctccacgc cgccgacgtt gccagcgcgc
     2041 gcgggtgcca ccgccagccc aggtccggcc ccagctgcct ccagggcgcg tcggcttggg
     2101 gcccagcgca ggtcagcgcc cgcgtcaaag taggactcgg cctctctatc gccgctgccc
     2161 gtgccagcca gggccctttg caggctgtgc atcagctcgc ggtcgctgag ctcgcgccgc
     2221 cggctcacgc tcacggcctt gtggatgcgc tcgttgcgat aaacgcccag gtcgtcgctc
     2281 aaggtaagca ccttcagcgc catgcgcatg tagaacccct cgatctttac ctccttgtct
     2341 atgggaacgt aaggggtatg gtatatcttg cgggcgtaaa acttgcccag gctaagcatg
     2401 gaatagttga tggcggccac cttgtcagcc aggctcaagc tgcgctcctg caccactatg
     2461 ctctgcagga tgtttatcaa atcgagcagc cagcggccct cgggctctac tatgtttagc
     2521 agcgcatccc tgaatgcctc gttgtccctg ctgtgctgca ctataaggaa cagctgcgcc
     2581 atgagcggct tgctatttgg gttttgctcc agcgcgctta caaagtccca cagatgcatc
     2641 agtcctatag ccacctcctc gcgcgccaca agcgtacgca cgtggttgtt aaagcttttt
     2701 tgaaagttaa tctcctggtt caccgtctgc tcgtatgcgg ttaccaggtc ggcggccgcc
     2761 acgtgtgcgc gcgcgggact aatcccggtt cgcgcgtcgg gctcaaagtc ctcctcgcgc
     2821 agcaaccgct cgcgattcag gccatgccgc agctcgcgcc ctgcgtggaa ctttcgatcc
     2881 cgcatctcct cgggctcctc tccctcgcgg tcgcgaaaca ggttctgccg cggcacgtac
     2941 gcctcacgcg tatcacgctt cagctgcacc cttgggtgcc gctcaggaga gggcgctcct
     3001 agccgcgcca ggccctcgcc ctcctccaag tccaggtagt gccgggcccg gcgccgcggg
     3061 ggttcgtaat caccatctgc tgccgcgtca accgcggatg tcgcccctcc tgacgcggta
     3121 ggaggagggg agggtgccct gcatgtctgc cgctgctctt gctcttgccg ctgctgagga
     3181 ggggggcgca tctgccgcag caccggatgc atctcgagat ctgagtccgg tagcgctagc
     3241 ggatctgacg gttcacacta gcggatctga cggttcacgc tagcggatct gacggttcac
     3301 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     3361 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     3421 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     3481 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     3541 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3601 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3661 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3721 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3781 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3841 taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata cggttatcca
     3901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg
     4741 ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg
     4801 gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg
     4861 gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     4921 caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg
     4981 tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat
     5041 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     5101 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     5161 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     5221 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     5281 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     5341 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     5401 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag
     5461 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     5521 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     5581 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     5641 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     5701 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     5761 cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga
     5821 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     5881 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     5941 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     6001 ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     6061 atagctcaga ggccgaggcg gcctcggcct ctgcataaat
//
