LOCUS       sequence-360303-        9367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible Salmonella Typhimurium SopB for expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9367)
  AUTHORS   Walpole GFW, Pacheco J, Chauhan N, Clark J, Anderson KE, Abbas YM, 
            Brabant-Kirwan D, Montano-Rendon F, Liu Z, Zhu H, Brumell JH, 
            Deiters A, Stephens LR, Hawkins PT, Hammond GRV, Grinstein S, Fairn 
            GD
  TITLE     Kinase-independent synthesis of 3-phosphorylated phosphoinositides 
            by a phosphotransferase.
  JOURNAL   Nat Cell Biol. 2022 Apr 28. pii: 10.1038/s41556-022-00895-y. doi: 
            10.1038/s41556-022-00895-y.
  PUBMED    35484249
REFERENCE   2  (bases 1 to 9367)
  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"; journalName: "Nat Cell 
            Biol. 2022 Apr 28. pii: 10.1038/s41556-022-00895-y. doi: 
            10.1038/s41556-022-00895-y."
COMMENT     Sequence Label: pESC-LEU-SopB
FEATURES             Location/Qualifiers
     source          1..9367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..59)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     159..178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(672..1766)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1767..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2475..2930)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2612..2633)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2824..2843
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3009..3174)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     3095..3119
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3322..3345)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        3392..4056
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3608..3725
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4020..4043
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      5782..5971
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6043..6060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6214..6802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6294..6313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6973..7764,7765..7833))
                     /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     7596..7615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7834..7938)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7965..9307
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(8928..8975)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     9348..9366
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
       61 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      121 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      181 gagagtgcac catatcgact acgtcgttaa ggccgtttct gacagagtaa aattcttgag
      241 ggaactttca ccattatggg aaatggttca agaaggtatt gacttaaact ccatcaaatg
      301 gtcaggtcat tgagtgtttt ttatttgttg tatttttttt tttttagaga aaatcctcca
      361 atatataaat taggaatcat agtttcatga ttttctgtta cacctaactt tttgtgtggt
      421 gccctcctcc ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc
      481 cattagtatc aatttgctta cctgtattcc tttacatcct cctttttctc cttcttgata
      541 aatgtatgta gattgcgtat atagtttcgt ctaccctatg aacatattcc attttgtaat
      601 ttcgtgtcgt ttctattatg aatttcattt ataaagttta tgtacaaata tcataaaaaa
      661 agagaatctt tttaagcaag gattttctta acttcttcgg cgacagcatc accgacttcg
      721 gtggtactgt tggaaccacc taaatcacca gttctgatac ctgcatccaa aaccttttta
      781 actgcatctt caatggcctt accttcttca ggcaagttca atgacaattt caacatcatt
      841 gcagcagaca agatagtggc gatagggttg accttattct ttggcaaatc tggagcagaa
      901 ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt ctggcaaaga ggccaaggac
      961 gcagatggca acaaacccaa ggaacctggg ataacggagg cttcatcgga gatgatatca
     1021 ccaaacatgt tgctggtgat tataatacca tttaggtggg ttgggttctt aactaggatc
     1081 atggcggcag aatcaatcaa ttgatgttga accttcaatg tagggaattc gttcttgatg
     1141 gtttcctcca cagtttttct ccataatctt gaagaggcca aaacattagc tttatccaag
     1201 gaccaaatag gcaatggtgg ctcatgttgt agggccatga aagcggccat tcttgtgatt
     1261 ctttgcactt ctggaacggt gtattgttca ctatcccaag cgacaccatc accatcgtct
     1321 tcctttctct taccaaagta aatacctccc actaattctc tgacaacaac gaagtcagta
     1381 cctttagcaa attgtggctt gattggagat aagtctaaaa gagagtcgga tgcaaagtta
     1441 catggtctta agttggcgta caattgaagt tctttacgga tttttagtaa accttgttca
     1501 ggtctaacac taccggtacc ccatttagga ccacccacag cacctaacaa aacggcatca
     1561 gccttcttgg aggcttccag cgcctcatct ggaagtggaa cacctgtagc atcgatagca
     1621 gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat tggaacgaac atcagaaata
     1681 gctttaagaa ccttaatggc ttcggctgtg atttcttgac caacgtggtc acctggcaaa
     1741 acgacgatct tcttaggggc agacattaga atggtatatc cttgaaatat atatatatat
     1801 attgctgaaa tgtaaaaggt aagaaaagtt agaaagtaag acgattgcta accacctatt
     1861 ggaaaaaaca ataggtcctt aaataatatt gtcaacttca agtattgtga tgcaagcatt
     1921 tagtcatgaa cgcttctcta ttctatatga aaagccggtt ccggcgctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt agatatacta
     2401 gaagttctcc tcgaccggtc gatatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2461 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2521 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2581 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2641 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2701 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2761 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2821 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2881 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     2941 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3001 ctgaattgga gcgacctcat gctatacctg agaaagcaac ctgacctaca ggaaagagtt
     3061 actcaagaat aagaattttc gttttaaaac ctaagagtca ctttaaaatt tgtatacact
     3121 tatttttttt ataacttatt taataataaa aatcataaat cataagaaat tcgcttattt
     3181 agaagtgtca acaacgtatc taccaacgat ttgacccttt tccatctttt cgtaaatttc
     3241 tggcaaggta gacaagccga caaccttgat tggagacttg accaaacctc tggcgaagaa
     3301 ttgttaatta agagctcaga tcttatcgtc gtcatccttg taatccatcg atactagtgc
     3361 ggccgccctt tagtgagggt tgaattcgaa ttttcaaaaa ttcttacttt ttttttggat
     3421 ggacgcaaag aagtttaata atcatattac atggcattac caccatatac atatccatat
     3481 acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta
     3541 gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat
     3601 tgaagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
     3661 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     3721 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3781 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
     3841 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3901 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
     3961 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
     4021 atatacctct atactttaac gtcaaggaga aaaaaccccg gatccaatgc aaatacagag
     4081 cttctatcac tcagcttcac taaaaaccca ggaggctttt aaaagcctac aaaaaacctt
     4141 atacaacgga atgcagattc tctcaggcca gggcaaagcg ccggctaaag cgcccgacgc
     4201 tcgcccggaa attattgtcc tgcgagaacc cggcgcgaca tgggggaatt atctacagca
     4261 tcagaaggcg tctaaccact cgctgcataa cctctataac ttacagcgcg atcttcttac
     4321 cgtcgcggca accgttctgg gtaaacaaga cccggttcta acgtcaatgg caaaccaaat
     4381 ggagttagcc aaagttaaag cggaccggcc agcaacaaaa caagaagaag ccgcggcaaa
     4441 agcattgaag aaaaatctta tcgaacttat tgcagcacgc actcagcagc aggatggctt
     4501 acctgcaaaa gaagctcatc gctttgcggc agtagcgttt agagatgctc aggtcaagca
     4561 gcttaataac cagccctggc aaaccataaa aaatacactc acgcataacg ggcatcacta
     4621 taccaacacg cagctccctg cagcagagat gaaaatcggc gcaaaagata tctttcccag
     4681 tgcttatgag ggaaagggcg tatgcagttg ggataccaag aatattcatc acgccaataa
     4741 tttgtggatg tccacggtga gtgtgcatga ggacggtaaa gataaaacgc ttttttgcgg
     4801 gatacgtcat ggcgtgcttt ccccctatca tgaaaaagat ccgcttctgc gtcacgtcgg
     4861 cgctgaaaac aaagccaaag aagtattaac tgcggcactt tttagtaaac ctgagttgct
     4921 taacaaagcc ttagcgggcg aggcggtaag cctgaaactg gtatccgtcg ggttactcac
     4981 cgcgtcgaat attttcggca aagagggaac gatggtcgag gaccaaatgc gcgcatggca
     5041 atcgttgacc cagccgggaa aaatgattca tttaaaaatc cgcaataaag atggcgatct
     5101 acagacggta aaaataaaac cggacgtcgc cgcatttaat gtgggtgtta atgagctggc
     5161 gctcaagctc ggctttggcc ttaaggcatc ggatagctat aatgccgagg cgctacatca
     5221 gttattaggc aatgatttac gccctgaagc cagaccaggt ggctgggttg gcgaatggct
     5281 ggcgcaatac ccggataatt atgaggtcgt caatacatta gcgcgccaga ttaaggatat
     5341 atggaaaaat aaccaacatc ataaagatgg cggcgaaccc tataaactcg cacaacgcct
     5401 tgccatgtta gcccatgaaa ttgacgcggt acccgcctgg aattgtaaaa gcggcaaaga
     5461 tcgtacaggg atgatggatt cagaaatcaa gcgagagatc atttccttac atcagaccca
     5521 tatgttaagt gcgcctggta gtcttccgga tagcggtgga cagaaaattt tccaaaaagt
     5581 attactgaat agcggtaacc tggagattca gaaacaaaat acgggcgggg cgggaaacaa
     5641 agtaatgaaa aatttatcgc cagaggtgct caatctttcc tatcaaaaac gagttgggga
     5701 tgaaaatatt tggcagtcag taaaaggcat ttcttcatta atcacatctc tcgagtaagc
     5761 ttggtaccgc ggctagctaa gatccgctct aaccgaaaag gaaggagtta gacaacctga
     5821 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     5881 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     5941 cttgcttgag aaggttttgg gacgctcgaa gatccagctg cattaatgaa tcggccaacg
     6001 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa gcatctgtgc
     7981 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     8041 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     8101 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     8161 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     8221 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     8281 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     8341 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     8401 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     8461 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     8521 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     8581 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     8641 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     8701 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     8761 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     8821 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     8881 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     8941 tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt
     9001 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg
     9061 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa
     9121 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata
     9181 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta
     9241 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata
     9301 ttggatcata ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     9361 cgaggcc
//
