LOCUS       sequence-366655-        7601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged triple RGG scaffold with N-terminal TsCC(A) 
            coiled coil. Expression induced in media containing galactose..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7601)
  AUTHORS   Garabedian MV, Wang W, Dabdoub JB, Tong M, Caldwell RM, Benman W, 
            Schuster BS, Deiters A, Good MC
  TITLE     Designer membraneless organelles sequester native factors for 
            control of cell behavior.
  JOURNAL   Nat Chem Biol. 2021 Sep;17(9):998-1007. doi: 
            10.1038/s41589-021-00840-4. Epub 2021 Aug 2.
  PUBMED    34341589
REFERENCE   2  (bases 1 to 7601)
  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"; doi: 
            "10.1038/s41589-021-00840-4"; journalName: "Nat Chem Biol"; date: 
            "2021-09"; volume: "17"; issue: "9"; pages: "998-1007"
COMMENT     Sequence Label: Yiplac211 proGAL1-TsCC(A)-RGG-GFP-RGG-RGG
FEATURES             Location/Qualifiers
     source          1..7601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        243..684
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    243..360
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     655..678
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2140..2160
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2161..2163,2164..2166,2167..2877)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2206..2227)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2467..2486)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2814..2835
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3900..4087
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3955..3979)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4099..4116)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4099..4115)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4108..4130)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4324..4343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4443..4465
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4583..5386)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5387..5602)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5609..5627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5695..5799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5800..5868,5869..6660)
                     /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(6018..6037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6831..7419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7320..7339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7573..7590
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta
      241 gacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct cctccgtgcg
      301 tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg
      361 aacaataaag attctacaat actagctttt atggttatga agaggaaaaa ttggcagtaa
      421 cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga tgataatgcg
      481 attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta
      541 ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt tcaacatttt
      601 cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat tgttaatata
      661 cctctatact ttaacgtcaa ggaggagctc atggtggaag tggctgaaga agtgaaggcc
      721 gtctccgccg cgctgtccga acgcatcacc cagctggcga cagaactgaa tgacaaggcg
      781 gtccgggctg cagaacgccg ggttgcggaa gtcacgcgtg ctgccggtga acagaccgca
      841 caggcagagc gggagctggc cgacgccgcg cagacagtcg acgacctgga agaaaaactg
      901 gttgaactgc aggacagata tgacagtttg acgctggcgc tggagtcaga acgttcactg
      961 cgtcagcagc atgatgtgga gatggcccag ctgaaagagc gtcttgcggc cgctgaagag
     1021 aatacccgtc agcgacgtga acggtatcag gagcagaaga cagtgctgca ggatgcgctt
     1081 aatgcggagc aggcacagca caaaaacacg cgggaagacc tgcagaaacg actggagcaa
     1141 atttctgtcg aagctaatgc gcgtacagaa gaactgaagt ctgaacgcga taaagtcaat
     1201 actttcctta cccgccttga atcgcaggaa aatgcgctgg cctcacgtcg tcagcagcat
     1261 ctggccaccc gcgaaacgct gcagcaacgc ctcgagcagg ccatcgctga cacgcaggcg
     1321 cgcgccggtg agattgcact tgaacgtgac agagtcagca gcctcaccgc aaggctggaa
     1381 tcgcaggaaa aggcctcctc ggagcaactg gtgcgtatgg gcagtgaaat agccagtctg
     1441 acagagcgtt gcacacagct ggaaaaccag cgtgatgatg cccgtctgga gacgatgggg
     1501 gagaaagaaa cggtcgcggc actgcgtggt gaggctgaag ccctgaagcg tcagaaccag
     1561 tcactgatgg cggcgctttc aggcaataaa cagaccggtg gccagaatgc gggatccatg
     1621 gaatcgaatc aatctaacaa cggtggatcg ggaaacgcag cattgaatcg tgggggccgt
     1681 tacgtaccac cacacttgag aggcggagat ggtggtgccg ccgctgctgc ctcagccgga
     1741 ggcgacgaca gacgcggtgg ggccgggggt ggtggctatc gtagaggtgg tggaaattca
     1801 gggggcggcg gcggaggagg ctacgatcgt ggatacaacg acaaccggga tgaccgtgac
     1861 aatcggggcg gctccggcgg ttatggccgg gacagaaatt acgaggatcg cggttataac
     1921 gggggtggtg ggggaggagg caatcgcggc tacaataata accgcggtgg gggtggaggc
     1981 ggctataatc ggcaggaccg tggagacggc ggttctagta atttctcgcg gggagggtat
     2041 aacaacagag atgagggctc agacaacaga ggctcaggca gatcatacaa caacgatcgg
     2101 cgcgacaatg gtggtgacgg gggcggaggc ggttccctcg agaatttata tttccagggc
     2161 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2221 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2281 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2341 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2401 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2461 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2521 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2581 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2641 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2701 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2761 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2821 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggaa
     2881 tctaaccaat caaacaatgg cggctccggt aacgcggcgc ttaacagggg cggtcgttac
     2941 gtgcctccgc acctgagagg gggcgacgga ggtgcggctg ctgccgcgag tgccggtggt
     3001 gatgacaggc gtggtggtgc gggtggcggg ggctatcgtc gtggaggcgg taattccggg
     3061 ggaggtggtg gtgggggtta cgacagaggg tacaatgata accgtgatga tcgtgacaac
     3121 agaggcggga gtggcggtta cggtagggat cgtaactatg aagacagagg ctataacggg
     3181 ggtggtggtg gcggtggcaa tagaggttat aataacaacc gtggcggcgg tggaggcggg
     3241 tataacagac aggatagagg ggacggcggc agtagcaact tctcccgtgg cggatataac
     3301 aaccgtgacg aaggctctga caataggggt tctggtaggt cttacaataa tgataggaga
     3361 gacaatggcg gcgacggtaa gcttatggaa tccaatcaat ccaataacgg tggctcgggt
     3421 aatgccgctc tgaatcgtgg tggccgctac gtgccgccgc atctgcgtgg tggcgatggt
     3481 ggtgcagcag cagctgcatc tgctggcggt gatgaccgtc gcggcggtgc gggcggtggc
     3541 ggttatcgtc gcggcggtgg caatagtggt ggcggtggcg gtggcggtta tgatcgtggt
     3601 tacaatgaca accgtgatga ccgcgataac cgtggcggtt ccggcggtta cggtcgtgat
     3661 cgtaattatg aagaccgtgg ttacaacggc ggtggcggtg gcggtggcaa tcgtggctat
     3721 aacaataacc gtggtggcgg tggcggtggc tacaatcgcc aggatcgtgg tgacggtggc
     3781 agctctaact tttcgcgcgg tggctataat aaccgtgatg aaggcagcga caaccgcggt
     3841 agtggccgtt cctacaataa cgatcgtcgc gacaatggtg gcgatggcta atgaccggtg
     3901 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     3961 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     4021 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     4081 ctaccggctg cagaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4141 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4201 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     4261 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     4321 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4381 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4441 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4501 gggcgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     4561 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     4621 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     4681 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     4741 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     4801 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     4861 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     4921 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     4981 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     5041 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     5101 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     5161 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     5221 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     5281 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     5341 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     5401 tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     5461 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     5521 tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat
     5581 aaaaaaaaaa tgatgaattg aaaagctggc ctcgtgatac gcctattttt ataggttaat
     5641 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5701 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5761 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5821 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5881 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5941 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6001 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6061 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6121 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6181 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6241 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6301 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6361 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6421 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6481 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6541 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6601 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6661 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6721 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6781 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6841 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6901 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6961 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7021 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7081 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7141 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7201 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7261 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7321 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7381 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7441 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7501 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7561 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a
//
