LOCUS       sequence-366531-        5841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for cloning PhoCl fused to TsCC(B) into yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5841)
  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 5841)
  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 PhoCl-TsCC(B)::KANMX
FEATURES             Location/Qualifiers
     source          1..5841
                     /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             256..981
                     /codon_start=1
                     /product="photocleavable protein that separates into two 
                     fragments upon exposure to violet light (Zhang et al., 
                     2017)"
                     /label=PhoCl
                     /translation="VIPDYFKQSFPEGYSWERSMTYEDGGICIATNDITMEGDSFINKI
                     HFKGTNFPPNGPVMQKRTVGWEASTEKMYERDGVLKGDVKMKLLLKGGGHYRCDYRTTY
                     KVKQKPVKLPDYHFVDHRIEILSHDKDYNKVKLYEHAVARNSTDSMDELYKGGSGGMVS
                     KGEETITSVIKPDMKNKLRMEGNVNGHAFVIEGEGSGKPFEGIQTIDLEVKEGAPLPFA
                     YDILTTAFHYGNRVFTKYPR"
     terminator      1938..2125
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1993..2017)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2200..3556
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2200..2543
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2544..3353
                     /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(2611..2630)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3359..3556
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3660..3679)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3661..3679)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3766..3783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3937..4525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4017..4036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4696..5487,5488..5556))
                     /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     5319..5338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5557..5661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5729..5747
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5785..5807)
                     /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 aaatggtgat ccctgactac ttcaagcaga gcttccccga gggctacagc
      301 tgggagcgca gcatgaccta cgaggacggc ggcatctgca tcgccaccaa cgacatcaca
      361 atggaggggg acagcttcat caacaagatc cacttcaagg gcacgaactt cccccccaac
      421 ggccccgtga tgcagaagag gaccgtgggc tgggaggcca gcaccgagaa gatgtacgag
      481 cgcgacggcg tgctgaaggg cgacgtgaag atgaagctgc tgctgaaggg cggcggccac
      541 tatcgctgcg actaccgcac cacctacaag gtcaagcaga agcccgtaaa gctgcccgac
      601 taccacttcg tggaccaccg catcgagatc ctgagccacg acaaggacta caacaaggtg
      661 aagctgtacg agcacgccgt ggcccgcaac tccaccgaca gcatggacga gctgtacaag
      721 ggtggcagcg gtggcatggt gagcaagggc gaggagacca ttacaagcgt gatcaagcct
      781 gacatgaaga acaagctgcg catggagggc aacgtgaacg gccacgcctt cgtgatcgag
      841 ggcgagggca gcggcaagcc cttcgagggc atccagacga ttgatttgga ggtgaaggag
      901 ggcgccccgc tgcccttcgc ctacgacatc ctgaccaccg ccttccacta cggcaaccgc
      961 gtgttcacca agtacccacg gtccggtggg ggaggggtgg aagtggctga agaagtgaag
     1021 gccgtctccg ccgcgctgtc cgaacgcatc acccagctgg cgacagaact gaatgacaag
     1081 gcggtccggg ctgcagaacg ccgggttgcg gaagtcacgc gtgctgccgg tgaacagacc
     1141 gcacaggcag agcgggagct ggccgacgcc gcgcagacag tcgacgacct ggaagaaaaa
     1201 ctggttgaac tgcaggacag atatgacagt ttgacgctgg cgctggagtc agaacgttca
     1261 ctgcgtcagc agcatgatgt ggagatggcc cagctgaaag agcgtcttgc ggccgctgaa
     1321 gagaataccc gtcaggaaga ggaacggtat caggagcaga agacagtgct gcaggatgcg
     1381 cttaatgcgg agcaggcaca gcacaaaaac acgcgggaag acctgcagaa acgactggag
     1441 caaatttctg tcgaagctaa tgcgcgtaca gaagaactga agtctgaacg cgataaagtc
     1501 aatactttcc ttacccgcct tgaatcgcag gaaaatgcgc tggcctcaga agaacagcag
     1561 catctggcca cccgcgaaac gctgcagcaa cgcctcgagc aggccatcgc tgacacgcag
     1621 gcgcgcgccg gtgagattgc acttgaacgt gacagagtca gcagcctcac cgcaaggctg
     1681 gaatcgcagg aaaaggcctc ctcggagcaa ctggtgcgta tgggcagtga aatagccagt
     1741 ctgacagagc gttgcacaca gctggaaaac cagcgtgatg atgcccgtct ggagacgatg
     1801 ggggagaaag aaacggtcgc ggcactgcgt ggtgaggctg aagccctgaa gcgtcagaac
     1861 cagtcactga tggcggcgct ttcaggcaat aaacagaccg gtggccagaa tgcgtaaggc
     1921 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     1981 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     2041 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     2101 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
     2161 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     2221 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     2281 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     2341 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     2401 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     2461 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     2521 atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat
     2581 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     2641 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     2701 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     2761 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2821 ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg
     2881 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2941 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     3001 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     3061 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
     3121 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     3181 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     3241 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     3301 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac
     3361 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     3421 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     3481 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     3541 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc
     3601 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     3661 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     3721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5701 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5761 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5821 ttgtctgtaa gcggatgccg g
//
