LOCUS       sequence-471772-        6171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of 52K-GFP-KS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6171)
  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 6171)
  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
FEATURES             Location/Qualifiers
     source          1..6171
                     /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(1271..1981,1982..1984))
                     /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     1662..1681
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1921..1942
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3378..3581)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3407..3427)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3582..3885
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4977..5024
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5049..5068
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5256..6050)
                     /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(5256..6050)
                     /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(5367..5386)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5977..5996
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6085..6171,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6099..6171,1..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 gacaggagaa gaagaactgc
     1141 gctagaacca acttattcat catcatcatc ttgtacagct cgtccatgcc gagagtgagt
     1201 cgcggccgct ttacttctat agacaggaga agaagaactg cgctagaacc aacttattca
     1261 tcatcatcat cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     1321 ccttcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc
     1381 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     1441 agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct
     1501 tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact
     1561 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     1621 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     1681 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     1741 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
     1801 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
     1861 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     1921 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     1981 tcactatggt gacgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt
     2041 actcgccgtc ctctggctcg tactcatcgt cctcgtcata ttcctccacg ccgccgacgt
     2101 tgccagcgcg cgcgggtgcc accgccagcc caggtccggc cccagctgcc tccagggcgc
     2161 gtcggcttgg ggcccagcgc aggtcagcgc ccgcgtcaaa gtaggactcg gcctctctat
     2221 cgccgctgcc cgtgccagcc agggcccttt gcaggctgtg catcagctcg cggtcgctga
     2281 gctcgcgccg ccggctcacg ctcacggcct tgtggatgcg ctcgttgcga taaacgccca
     2341 ggtcgtcgct caaggtaagc accttcagcg ccatgcgcat gtagaacccc tcgatcttta
     2401 cctccttgtc tatgggaacg taaggggtat ggtatatctt gcgggcgtaa aacttgccca
     2461 ggctaagcat ggaatagttg atggcggcca ccttgtcagc caggctcaag ctgcgctcct
     2521 gcaccactat gctctgcagg atgtttatca aatcgagcag ccagcggccc tcgggctcta
     2581 ctatgtttag cagcgcatcc ctgaatgcct cgttgtccct gctgtgctgc actataagga
     2641 acagctgcgc catgagcggc ttgctatttg ggttttgctc cagcgcgctt acaaagtccc
     2701 acagatgcat cagtcctata gccacctcct cgcgcgccac aagcgtacgc acgtggttgt
     2761 taaagctttt ttgaaagtta atctcctggt tcaccgtctg ctcgtatgcg gttaccaggt
     2821 cggcggccgc cacgtgtgcg cgcgcgggac taatcccggt tcgcgcgtcg ggctcaaagt
     2881 cctcctcgcg cagcaaccgc tcgcgattca ggccatgccg cagctcgcgc cctgcgtgga
     2941 actttcgatc ccgcatctcc tcgggctcct ctccctcgcg gtcgcgaaac aggttctgcc
     3001 gcggcacgta cgcctcacgc gtatcacgct tcagctgcac ccttgggtgc cgctcaggag
     3061 agggcgctcc tagccgcgcc aggccctcgc cctcctccaa gtccaggtag tgccgggccc
     3121 ggcgccgcgg gggttcgtaa tcaccatctg ctgccgcgtc aaccgcggat gtcgcccctc
     3181 ctgacgcggt aggaggaggg gagggtgccc tgcatgtctg ccgctgctct tgctcttgcc
     3241 gctgctgagg aggggggcgc atctgccgca gcaccggatg catctcgaga tctgagtccg
     3301 gtagcgctag cggatctgac ggttcacact agcggatctg acggttcacg ctagcggatc
     3361 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
     3421 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     3481 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     3541 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     3601 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     3661 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     3721 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     3781 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     3841 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     3901 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat
     3961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4801 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     4861 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     4921 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     4981 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     5041 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     5101 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     5161 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     5221 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     5281 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     5341 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     5401 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     5461 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     5521 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     5581 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     5641 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     5701 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     5761 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     5821 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     5881 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     5941 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     6001 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     6061 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     6121 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa t
//
