LOCUS       sequence-471757-        5261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of 52K-KS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5261)
  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 5261)
  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
FEATURES             Location/Qualifiers
     source          1..5261
                     /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(2468..2671)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2497..2517)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2672..2975
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3150..3738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3230..3249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4067..4114
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4139..4158
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4346..5140)
                     /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(4346..5140)
                     /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(4457..4476)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5067..5086
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5175..5261,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5189..5261,1..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 tgtcgaccta
     1141 ctatagacag gagaagaaga actgcgctag aaccaactta ttcatcatca tcatgtactc
     1201 gccgtcctct ggctcgtact catcgtcctc gtcatattcc tccacgccgc cgacgttgcc
     1261 agcgcgcgcg ggtgccaccg ccagcccagg tccggcccca gctgcctcca gggcgcgtcg
     1321 gcttggggcc cagcgcaggt cagcgcccgc gtcaaagtag gactcggcct ctctatcgcc
     1381 gctgcccgtg ccagccaggg ccctttgcag gctgtgcatc agctcgcggt cgctgagctc
     1441 gcgccgccgg ctcacgctca cggccttgtg gatgcgctcg ttgcgataaa cgcccaggtc
     1501 gtcgctcaag gtaagcacct tcagcgccat gcgcatgtag aacccctcga tctttacctc
     1561 cttgtctatg ggaacgtaag gggtatggta tatcttgcgg gcgtaaaact tgcccaggct
     1621 aagcatggaa tagttgatgg cggccacctt gtcagccagg ctcaagctgc gctcctgcac
     1681 cactatgctc tgcaggatgt ttatcaaatc gagcagccag cggccctcgg gctctactat
     1741 gtttagcagc gcatccctga atgcctcgtt gtccctgctg tgctgcacta taaggaacag
     1801 ctgcgccatg agcggcttgc tatttgggtt ttgctccagc gcgcttacaa agtcccacag
     1861 atgcatcagt cctatagcca cctcctcgcg cgccacaagc gtacgcacgt ggttgttaaa
     1921 gcttttttga aagttaatct cctggttcac cgtctgctcg tatgcggtta ccaggtcggc
     1981 ggccgccacg tgtgcgcgcg cgggactaat cccggttcgc gcgtcgggct caaagtcctc
     2041 ctcgcgcagc aaccgctcgc gattcaggcc atgccgcagc tcgcgccctg cgtggaactt
     2101 tcgatcccgc atctcctcgg gctcctctcc ctcgcggtcg cgaaacaggt tctgccgcgg
     2161 cacgtacgcc tcacgcgtat cacgcttcag ctgcaccctt gggtgccgct caggagaggg
     2221 cgctcctagc cgcgccaggc cctcgccctc ctccaagtcc aggtagtgcc gggcccggcg
     2281 ccgcgggggt tcgtaatcac catctgctgc cgcgtcaacc gcggatgtcg cccctcctga
     2341 cgcggtagga ggaggggagg gtgccctgca tgtctgccgc tgctcttgct cttgccgctg
     2401 ctgaggaggg gggcgcatct gccgcagcac cggatgcatg ctagcggatc tgacggttca
     2461 ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     2521 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     2581 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     2641 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     2701 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     2761 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     2821 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     2881 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     2941 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     3001 ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat acggttatcc
     3061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3781 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3841 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag
     3901 gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     3961 ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt
     4021 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
     4081 acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
     4141 gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga
     4201 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     4261 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     4321 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     4381 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     4441 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     4501 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     4561 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     4621 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     4681 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     4741 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4801 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4861 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4921 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     4981 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     5041 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     5101 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     5161 tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5221 aatagctcag aggccgaggc ggcctcggcc tctgcataaa t
//
