LOCUS       sequence-366596-        5817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for cloning mScarlet fused to TsCC(B) coiled coil into 
            yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5817)
  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 5817)
  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: pfa6a mScarlet-TsCC(B)::KANMX
FEATURES             Location/Qualifiers
     source          1..5817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..85
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        166..184
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     166..183
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             946..966
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1914..2101
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1969..1993)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2176..3532
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2176..2519
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2520..3329
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2587..2606)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3335..3532
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3636..3655)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3637..3655)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3742..3759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3913..4501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3993..4012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4672..5463,5464..5532))
                     /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     5295..5314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5533..5637)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5705..5723
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5761..5783)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
       61 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga
      121 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
      181 tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcg gtgacggtgc
      241 tggtttaatt aagtatcaaa gggtgaggca gtcatcaaag agtttatgcg tttcaaagtt
      301 cacatggaag ggtccatgaa cgggcacgag ttcgagattg agggtgaggg ggagggaagg
      361 ccctacgagg gcacccagac agccaaattg aaagtaacga agggcggtcc tctgccgttt
      421 tcctgggata tattatcccc ccaattcatg tatggttcca gggccttcac caagcatccc
      481 gccgatatac ccgattatta caaacaatct ttcccagagg ggtttaagtg ggaaagagtt
      541 atgaatttcg aagacggagg tgcagtcacg gtcacccagg atacgtcatt agaggatggt
      601 acattaattt acaaggtcaa acttaggggc acaaatttcc cacctgatgg tcctgtgatg
      661 caaaagaaga ctatgggttg ggaggctagt acagagagat tgtacccgga ggatggagtc
      721 ttgaaagggg acataaaaat ggcactgagg ttgaaggacg gaggccgtta tctggcggac
      781 ttcaagacaa cctataaagc taagaagcct gtccagatgc ccggggcgta caatgtagat
      841 agaaaacttg acataaccag ccacaatgag gactatacag tagtcgaaca gtatgaaaga
      901 tctgaaggga ggcactcaac cggaggcatg gatgaactat acaaggagaa tctttatttc
      961 cagggactcg aggtggaagt ggctgaagaa gtgaaggccg tctccgccgc gctgtccgaa
     1021 cgcatcaccc agctggcgac agaactgaat gacaaggcgg tccgggctgc agaacgccgg
     1081 gttgcggaag tcacgcgtgc tgccggtgaa cagaccgcac aggcagagcg ggagctggcc
     1141 gacgccgcgc agacagtcga cgacctggaa gaaaaactgg ttgaactgca ggacagatat
     1201 gacagtttga cgctggcgct ggagtcagaa cgttcactgc gtcagcagca tgatgtggag
     1261 atggcccagc tgaaagagcg tcttgcggcc gctgaagaga atacccgtca ggaagaggaa
     1321 cggtatcagg agcagaagac agtgctgcag gatgcgctta atgcggagca ggcacagcac
     1381 aaaaacacgc gggaagacct gcagaaacga ctggagcaaa tttctgtcga agctaatgcg
     1441 cgtacagaag aactgaagtc tgaacgcgat aaagtcaata ctttccttac ccgccttgaa
     1501 tcgcaggaaa atgcgctggc ctcagaagaa cagcagcatc tggccacccg cgaaacgctg
     1561 cagcaacgcc tcgagcaggc catcgctgac acgcaggcgc gcgccggtga gattgcactt
     1621 gaacgtgaca gagtcagcag cctcaccgca aggctggaat cgcaggaaaa ggcctcctcg
     1681 gagcaactgg tgcgtatggg cagtgaaata gccagtctga cagagcgttg cacacagctg
     1741 gaaaaccagc gtgatgatgc ccgtctggag acgatggggg agaaagaaac ggtcgcggca
     1801 ctgcgtggtg aggctgaagc cctgaagcgt cagaaccagt cactgatggc ggcgctttca
     1861 ggcaataaac agaccggtgg ccagaatgcg taaggcgcgc cacttctaaa taagcgaatt
     1921 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     1981 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     2041 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg
     2101 gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg
     2161 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
     2221 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
     2281 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
     2341 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
     2401 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
     2461 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
     2521 tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt
     2581 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     2641 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2701 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2761 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     2821 gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2881 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2941 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3001 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3061 ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3121 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3181 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3241 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3301 ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt
     3361 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     3421 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     3481 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     3541 tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga tatcagatcc
     3601 actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc
     3661 gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3721 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4321 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4621 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4981 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5281 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5521 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5641 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5701 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5761 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgg
//
