LOCUS       sequence-471761-        5288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of 52K-linkerKS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5288)
  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 5288)
  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-linkerKS
FEATURES             Location/Qualifiers
     source          1..5288
                     /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(2495..2698)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2524..2544)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2699..3002
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3177..3765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3257..3276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4094..4141
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4166..4185
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4373..5167)
                     /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(4373..5167)
                     /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(4484..4503)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5094..5113
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5202..5288,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5216..5288,1..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 aagaagaact gcgctagaac caacttattc atcatcatca tagatcctcc
     1201 gcctccgctt cctccgccgc cgtactcgcc gtcctctggc tcgtactcat cgtcctcgtc
     1261 atattcctcc acgccgccga cgttgccagc gcgcgcgggt gccaccgcca gcccaggtcc
     1321 ggccccagct gcctccaggg cgcgtcggct tggggcccag cgcaggtcag cgcccgcgtc
     1381 aaagtaggac tcggcctctc tatcgccgct gcccgtgcca gccagggccc tttgcaggct
     1441 gtgcatcagc tcgcggtcgc tgagctcgcg ccgccggctc acgctcacgg ccttgtggat
     1501 gcgctcgttg cgataaacgc ccaggtcgtc gctcaaggta agcaccttca gcgccatgcg
     1561 catgtagaac ccctcgatct ttacctcctt gtctatggga acgtaagggg tatggtatat
     1621 cttgcgggcg taaaacttgc ccaggctaag catggaatag ttgatggcgg ccaccttgtc
     1681 agccaggctc aagctgcgct cctgcaccac tatgctctgc aggatgttta tcaaatcgag
     1741 cagccagcgg ccctcgggct ctactatgtt tagcagcgca tccctgaatg cctcgttgtc
     1801 cctgctgtgc tgcactataa ggaacagctg cgccatgagc ggcttgctat ttgggttttg
     1861 ctccagcgcg cttacaaagt cccacagatg catcagtcct atagccacct cctcgcgcgc
     1921 cacaagcgta cgcacgtggt tgttaaagct tttttgaaag ttaatctcct ggttcaccgt
     1981 ctgctcgtat gcggttacca ggtcggcggc cgccacgtgt gcgcgcgcgg gactaatccc
     2041 ggttcgcgcg tcgggctcaa agtcctcctc gcgcagcaac cgctcgcgat tcaggccatg
     2101 ccgcagctcg cgccctgcgt ggaactttcg atcccgcatc tcctcgggct cctctccctc
     2161 gcggtcgcga aacaggttct gccgcggcac gtacgcctca cgcgtatcac gcttcagctg
     2221 cacccttggg tgccgctcag gagagggcgc tcctagccgc gccaggccct cgccctcctc
     2281 caagtccagg tagtgccggg cccggcgccg cgggggttcg taatcaccat ctgctgccgc
     2341 gtcaaccgcg gatgtcgccc ctcctgacgc ggtaggagga ggggagggtg ccctgcatgt
     2401 ctgccgctgc tcttgctctt gccgctgctg aggagggggg cgcatctgcc gcagcaccgg
     2461 atgcatgcta gcggatctga cggttcacta aaccagctct gcttatatag acctcccacc
     2521 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     2581 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     2641 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     2701 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     2761 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     2821 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     2881 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     2941 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     3001 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     3061 taatgcatgg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3121 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3181 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3241 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3301 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3361 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3421 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3481 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3541 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3601 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3661 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3721 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3781 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3841 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3901 atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct
     3961 gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg
     4021 gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga
     4081 accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc
     4141 gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta
     4201 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     4261 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     4321 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     4381 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     4441 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     4501 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     4561 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     4621 cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     4681 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     4741 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     4801 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     4861 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     4921 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     4981 cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     5041 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     5101 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     5161 caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa gcctaggcct
     5221 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     5281 gcataaat
//
