LOCUS       sequence-151912-        5697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  create protein aggregates in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5697)
  AUTHORS   Miyazaki Y, Mizumoto K, Dey G, Kudo T, Perrino J, Chen LC, Meyer T, 
            Wandless TJ
  TITLE     A method to rapidly create protein aggregates in living cells.
  JOURNAL   Nat Commun. 2016 May 27;7:11689. doi: 10.1038/ncomms11689.
  PUBMED    27229621
REFERENCE   2  (bases 1 to 5697)
  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: "Nat 
            Commun."; date: "2016-05-27"; volume: "7:11689. doi"; pages: " 
            10.1038/ncomms11689"
COMMENT     Sequence Label: Pvha6-AgDD
FEATURES             Location/Qualifiers
     source          1..5697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1328..1648
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KPE"
     CDS             1655..2368
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1655..2365
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1682..1710)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2305..2325
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3216..3238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3338..3357
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3439..3457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3525..3629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3630..3698,3699..4490)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3848..3867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4661..5249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5150..5169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5403..5420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5537..5558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5573..5578,5579..5596,5597..5603)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5611..5627
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5616..5638
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgta cctttatagg tgcgctctac gcaattcacc
       61 agctgaacaa tggagttgag cctaatgtaa ctaaaaattt atttgaatgc tttacaaaaa
      121 tattatttca gatcttcgag atcatgaaaa ctatcaaaca gcagcgccct ggagcaatcg
      181 agtcgttccc acaatattca ggtgtatatg caatcgtttt agactacatt tcggtaagtt
      241 gctacttcag agataaactg taattatttt aaatttcagc gcaaacgcgg aggaaagtct
      301 gatcctgtta acaaatacat aaatcgtttc ctcgctgatc tcacggagat attgccagca
      361 tgctcaacgt tgccagtgat gaatccaagc acagaactgc attaagtata ctatttatta
      421 ctcgatactt ttgttcacat aggtttttta aatcatattt tatgcatcat ttatcatatt
      481 caatgcatca ttcatatcat agtcaataaa aaggttgatt tctcatgttc tggtttcaaa
      541 tgctgacttt ggtaaaaaga acgcgtgcct gcctattgcc tatcttggca ttttctcgat
      601 aaattttaaa atgtaggttc gatcttatga gatttgtagt caaaagagct catatgtatt
      661 caggtaggtc tggtagcgag accaacttaa tagcatgaca agcattttca atttgccctg
      721 gagcgcaatt ggttttttat tcgaaaatcg cacatttctg tttccccata atataaaatt
      781 tccaggacga tatatattac attcttcaca aaatattgca ttacagacac cgacaaagaa
      841 tctccacctg atatgaaaac aatgagccaa caatgttatc tgtattgcca ccacccacat
      901 ttcctagtca ttcagtatat attgtttcaa ttgaatcatt gcaggtatat atcgaattga
      961 acttgtaagg cttcatcttc atttctcaat acatcatcca tcattccaga gcagctccgg
     1021 ccacacaaaa attggtggcg gtctgatatt gataatcgac ttctttgacg tgcctgacgg
     1081 agcagcaaag cggagcactg ataagacaat gaagaactaa aaaattgtct tcggttttca
     1141 gtctttagtt ctgcagcact ttattttttg tttctcctat ttttccgcat tttcctaact
     1201 ttctgatgtc catttcaaat gatttttgtt ataaaattgt ttaatttcag ggcgactaaa
     1261 acctacccgg cgcgcctcta gaggatccgc caccatgctg gccctgaagc tggctggtct
     1321 agatattgga gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg
     1381 cggccagacc tgtgtggtgc actacaccgg gatgcttgaa gatggaaaga aagtcgattc
     1441 ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg
     1501 ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc
     1561 agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt
     1621 cttcgatgtg gagcttctaa aaccggaaga attcagcaaa ggagaagaac ttttcactgg
     1681 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtccg
     1741 tggagagggt gaaggtgatg ctacaaacgg aaaactcacc cttaaattta tttgcactac
     1801 tggaaaacta cctgttccgt ggccaacact tgtcactact ctgacctatg gtgttcaatg
     1861 cttttcccgt tatccggatc acatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     1921 aggttatgta caggaacgca ctatatcttt caaagatgac gggacctaca agacgcgtgc
     1981 tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaagg gtattgattt
     2041 taaagaagat ggaaacattc ttggacacaa actcgagtac aactttaact cacacaatgt
     2101 atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa
     2161 cgttgaagat ggttccgttc aactagcaga ccattatcaa caaaatactc caattggcga
     2221 tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctgtcc tttcgaaaga
     2281 tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac
     2341 acatggcatg gatgagctct acaaataagg taccatggta ttgatatctg agctccgcat
     2401 cggccgctgt catcagatcg ccatctcgcg cccgtgcctc tgacttctaa gtccaattac
     2461 tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt ttgttattat
     2521 caaaaaaact tcttcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     2581 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     2641 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     2701 cttatgtttt ttttacttct gataaatttt ttttgaaaca tcatagaaaa aaccgcacac
     2761 aaaatacctt atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg
     2821 tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg
     2881 aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttgggggt
     2941 ttcccctatt gtttgtcaag agtttcgagg acggcgtttt tcttgctaaa atcacaagta
     3001 ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct cagtggaagg
     3061 tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg ccttaaatga
     3121 cagaatacat tcccaatata ccaaacataa ctgtttccta ctagtcggcc gtacgggccc
     3181 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3241 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3301 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     3361 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     3421 aggcggcctt aattaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3481 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3541 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3601 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3661 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3721 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3781 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3841 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3901 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3961 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4021 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4081 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4141 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4201 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4261 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4321 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4381 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4441 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4501 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4561 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4621 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4681 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4741 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4801 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4861 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4921 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4981 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5041 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5101 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5161 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5221 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5281 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5341 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5401 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5461 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5521 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5581 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5641 acagctgggg gtgggggggg ggttgggggt ggggtgttgt tggggtgtgt tttgtgt
//
