LOCUS       sequence-366600-        6824 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GFP tagged triple RGG scaffold with N-terminal SynZip1 
            coiled coil (Sz1). Expression induced in media containing 
            galactose..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  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 6824)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SZ1-RGG-GFP-RGG-RGG
FEATURES             Location/Qualifiers
     source          1..6824
                     /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             1363..1383
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1384..1386,1387..1389,1390..2100)
                     /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(1429..1450)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1690..1709)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2037..2058
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3123..3310
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3178..3202)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3322..3339)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3322..3338)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3331..3353)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3547..3566)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3666..3688
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3806..4609)
                     /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(4610..4825)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4832..4850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5023..5091,5092..5883)
                     /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(5241..5260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6054..6642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6543..6562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6796..6813
                     /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 atgaacttgg tagcccagct tgagaacgaa
      721 gttgcgagtc tggaaaacga aaacgagacg ttgaagaaaa agaacttgca caagaaagat
      781 ttgattgcct acttggaaaa ggaaattgcc aatctgcgta aaaagattga ggaaggatcc
      841 atggaatcga atcaatctaa caacggtgga tcgggaaacg cagcattgaa tcgtgggggc
      901 cgttacgtac caccacactt gagaggcgga gatggtggtg ccgccgctgc tgcctcagcc
      961 ggaggcgacg acagacgcgg tggggccggg ggtggtggct atcgtagagg tggtggaaat
     1021 tcagggggcg gcggcggagg aggctacgat cgtggataca acgacaaccg ggatgaccgt
     1081 gacaatcggg gcggctccgg cggttatggc cgggacagaa attacgagga tcgcggttat
     1141 aacgggggtg gtgggggagg aggcaatcgc ggctacaata ataaccgcgg tgggggtgga
     1201 ggcggctata atcggcagga ccgtggagac ggcggttcta gtaatttctc gcggggaggg
     1261 tataacaaca gagatgaggg ctcagacaac agaggctcag gcagatcata caacaacgat
     1321 cggcgcgaca atggtggtga cgggggcgga ggcggttccc tcgagaattt atatttccag
     1381 ggcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1441 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1501 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1561 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1621 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1681 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1741 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1801 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1861 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1921 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1981 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     2041 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2101 gaatctaacc aatcaaacaa tggcggctcc ggtaacgcgg cgcttaacag gggcggtcgt
     2161 tacgtgcctc cgcacctgag agggggcgac ggaggtgcgg ctgctgccgc gagtgccggt
     2221 ggtgatgaca ggcgtggtgg tgcgggtggc gggggctatc gtcgtggagg cggtaattcc
     2281 gggggaggtg gtggtggggg ttacgacaga gggtacaatg ataaccgtga tgatcgtgac
     2341 aacagaggcg ggagtggcgg ttacggtagg gatcgtaact atgaagacag aggctataac
     2401 gggggtggtg gtggcggtgg caatagaggt tataataaca accgtggcgg cggtggaggc
     2461 gggtataaca gacaggatag aggggacggc ggcagtagca acttctcccg tggcggatat
     2521 aacaaccgtg acgaaggctc tgacaatagg ggttctggta ggtcttacaa taatgatagg
     2581 agagacaatg gcggcgacgg taagcttatg gaatccaatc aatccaataa cggtggctcg
     2641 ggtaatgccg ctctgaatcg tggtggccgc tacgtgccgc cgcatctgcg tggtggcgat
     2701 ggtggtgcag cagcagctgc atctgctggc ggtgatgacc gtcgcggcgg tgcgggcggt
     2761 ggcggttatc gtcgcggcgg tggcaatagt ggtggcggtg gcggtggcgg ttatgatcgt
     2821 ggttacaatg acaaccgtga tgaccgcgat aaccgtggcg gttccggcgg ttacggtcgt
     2881 gatcgtaatt atgaagaccg tggttacaac ggcggtggcg gtggcggtgg caatcgtggc
     2941 tataacaata accgtggtgg cggtggcggt ggctacaatc gccaggatcg tggtgacggt
     3001 ggcagctcta acttttcgcg cggtggctat aataaccgtg atgaaggcag cgacaaccgc
     3061 ggtagtggcc gttcctacaa taacgatcgt cgcgacaatg gtggcgatgg ctaatgaccg
     3121 gtgcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     3181 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     3241 ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca
     3301 cctctaccgg ctgcagaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     3361 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     3421 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     3481 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     3541 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3601 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3661 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3721 aaagggcggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     3781 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     3841 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     3901 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     3961 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     4021 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     4081 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     4141 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     4201 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     4261 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     4321 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     4381 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     4441 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     4501 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     4561 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     4621 gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     4681 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     4741 ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag
     4801 aataaaaaaa aaatgatgaa ttgaaaagct ggcctcgtga tacgcctatt tttataggtt
     4861 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5701 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6721 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6781 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
