LOCUS       sequence-366492-        5226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for cloning mScarlet fused to FKBP into yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5226)
  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 5226)
  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-FKBP::KANMX
FEATURES             Location/Qualifiers
     source          1..5226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             764..784
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(791..793,794..796,797..1120)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MKGVQVETISPGDGRTFHKRGQTCVVHYTGMLEDGKKFDSSRDRN
                     KPFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVE
                     LLKLE"
     terminator      1141..1328
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1196..1220)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1403..2759
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1403..1746
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1747..2556
                     /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(1814..1833)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2562..2759
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2863..2882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2864..2882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2969..2986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3140..3728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3220..3239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3899..4690,4691..4759))
                     /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     4522..4541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4760..4864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4932..4950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4988..5010)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5110..5129
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5210..5226,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5210..5226,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg
       61 gtttaattaa gtatcaaagg gtgaggcagt catcaaagag tttatgcgtt tcaaagttca
      121 catggaaggg tccatgaacg ggcacgagtt cgagattgag ggtgaggggg agggaaggcc
      181 ctacgagggc acccagacag ccaaattgaa agtaacgaag ggcggtcctc tgccgttttc
      241 ctgggatata ttatcccccc aattcatgta tggttccagg gccttcacca agcatcccgc
      301 cgatataccc gattattaca aacaatcttt cccagagggg tttaagtggg aaagagttat
      361 gaatttcgaa gacggaggtg cagtcacggt cacccaggat acgtcattag aggatggtac
      421 attaatttac aaggtcaaac ttaggggcac aaatttccca cctgatggtc ctgtgatgca
      481 aaagaagact atgggttggg aggctagtac agagagattg tacccggagg atggagtctt
      541 gaaaggggac ataaaaatgg cactgaggtt gaaggacgga ggccgttatc tggcggactt
      601 caagacaacc tataaagcta agaagcctgt ccagatgccc ggggcgtaca atgtagatag
      661 aaaacttgac ataaccagcc acaatgagga ctatacagta gtcgaacagt atgaaagatc
      721 tgaagggagg cactcaaccg gaggcatgga tgaactatac aaggagaatc tttatttcca
      781 gggactcgag atgaaaggcg tgcaggtgga gactatctcc ccaggagacg ggcgcacctt
      841 ccacaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa
      901 atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt
      961 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac
     1021 tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc
     1081 cactctcgtc ttcgatgtgg agcttctaaa actggaatag ggcgcgccac ttctaaataa
     1141 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     1201 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     1261 ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc
     1321 tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc ttgcctcgtc
     1381 cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     1441 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     1501 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     1561 ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     1621 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     1681 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     1741 acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc caacatggat
     1801 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1861 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1921 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1981 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2041 atccccggca aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2101 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2161 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2221 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2281 gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2341 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2401 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2461 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2521 ttgcagtttc atttgatgct cgatgagttt ttctaatcag tactgacaat aaaaagattc
     2581 ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat
     2641 caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt
     2701 taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc
     2761 tgtcgattcg atactaacgc cgccatccag tttaaacgag ctcgaattca tcgatgatat
     2821 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     2881 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2941 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3001 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3061 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4861 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4921 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4981 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5041 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     5101 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt
     5161 tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac
     5221 actata
//
