LOCUS       sequence-471793-        5258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of 52K-KS-FtoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5258)
  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 5258)
  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-KS-FtoA
FEATURES             Location/Qualifiers
     source          1..5258
                     /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"
     promoter        complement(2465..2668)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2494..2514)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2669..2972
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3147..3735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3227..3246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4064..4111
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4136..4155
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4343..5137)
                     /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(4343..5137)
                     /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(4454..4473)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5064..5083
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5172..5258,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5186..5258,1..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 atcagttatc tagatccggt ggatcccggg cccgcggtac cgtcgaccta
     1141 tagacaggag gcggcggcct gcgctagaac caacttattc atcatcatca tgtactcgcc
     1201 gtcctctggc tcgtactcat cgtcctcgtc atattcctcc acgccgccga cgttgccagc
     1261 gcgcgcgggt gccaccgcca gcccaggtcc ggccccagct gcctccaggg cgcgtcggct
     1321 tggggcccag cgcaggtcag cgcccgcgtc aaagtaggac tcggcctctc tatcgccgct
     1381 gcccgtgcca gccagggccc tttgcaggct gtgcatcagc tcgcggtcgc tgagctcgcg
     1441 ccgccggctc acgctcacgg ccttgtggat gcgctcgttg cgataaacgc ccaggtcgtc
     1501 gctcaaggta agcaccttca gcgccatgcg catgtagaac ccctcgatct ttacctcctt
     1561 gtctatggga acgtaagggg tatggtatat cttgcgggcg taaaacttgc ccaggctaag
     1621 catggaatag ttgatggcgg ccaccttgtc agccaggctc aagctgcgct cctgcaccac
     1681 tatgctctgc aggatgttta tcaaatcgag cagccagcgg ccctcgggct ctactatgtt
     1741 tagcagcgca tccctgaatg cctcgttgtc cctgctgtgc tgcactataa ggaacagctg
     1801 cgccatgagc ggcttgctat ttgggttttg ctccagcgcg cttacaaagt cccacagatg
     1861 catcagtcct atagccacct cctcgcgcgc cacaagcgta cgcacgtggt tgttaaagct
     1921 tttttgaaag ttaatctcct ggttcaccgt ctgctcgtat gcggttacca ggtcggcggc
     1981 cgccacgtgt gcgcgcgcgg gactaatccc ggttcgcgcg tcgggctcaa agtcctcctc
     2041 gcgcagcaac cgctcgcgat tcaggccatg ccgcagctcg cgccctgcgt ggaactttcg
     2101 atcccgcatc tcctcgggct cctctccctc gcggtcgcga aacaggttct gccgcggcac
     2161 gtacgcctca cgcgtatcac gcttcagctg cacccttggg tgccgctcag gagagggcgc
     2221 tcctagccgc gccaggccct cgccctcctc caagtccagg tagtgccggg cccggcgccg
     2281 cgggggttcg taatcaccat ctgctgccgc gtcaaccgcg gatgtcgccc ctcctgacgc
     2341 ggtaggagga ggggagggtg ccctgcatgt ctgccgctgc tcttgctctt gccgctgctg
     2401 aggagggggg cgcatctgcc gcagcaccgg atgcatgcta gcggatctga cggttcacta
     2461 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     2521 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     2581 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     2641 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     2701 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     2761 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     2821 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     2881 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     2941 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     3001 atgaccccgt aattgattac tattaataac taatgcatgg cggtaatacg gttatccaca
     3061 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3121 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3181 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3241 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3301 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3361 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3421 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3481 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3541 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3601 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3661 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3721 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3781 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3841 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     3901 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     3961 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     4021 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     4081 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     4141 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     4201 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     4261 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     4321 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4381 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4441 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4501 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4561 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4621 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     4681 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4741 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4801 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4861 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4921 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4981 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     5041 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5101 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     5161 cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     5221 agctcagagg ccgaggcggc ctcggcctct gcataaat
//
