LOCUS       sequence-360301-        9364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible Salmonella Typhimurium SopB (catalytically inactive) for 
            expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9364)
  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 9364)
  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(C460S)
FEATURES             Location/Qualifiers
     source          1..9364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      49..1388
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    392..425
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        1415..1519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1520..1588,1589..2380)
                     /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(1738..1757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2551..3139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3040..3059
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3293..3310
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3382..3571)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        5297..5961
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(5310..5333)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5628..5745
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             6008..6031
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      6179..6344
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6234..6258)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      6423..6878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6510..6529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6720..6741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7179..7586
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7587..8681
                     /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"
     primer_bind     complement(9175..9194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9294..9316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(join(9354..9364,1..8))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagtatga tccaatatca
       61 aaggaaatga tagcattgaa ggatgagact aatccaattg aggagtggca gcatatagaa
      121 cagctaaagg gtagtgctga aggaagcata cgataccccg catggaatgg gataatatca
      181 caggaggtac tagactacct ttcatcctac ataaatagac gcatataagt acgcatttaa
      241 gcataaacac gcactatgcc gttcttctca tgtatatata tacaggcaac acgcagatat
      301 aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc gttgcatttt cggaagcgct
      361 cgttttcgga aacgctttga agttcctatc cgaagttcct attctctaga aagtatagga
      421 acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa
      481 acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct
      541 caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct acccatccac
      601 ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag
      661 tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa
      721 tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc
      781 ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg
      841 caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc
      901 agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa
      961 aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca
     1021 agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga
     1081 aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg
     1141 gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt
     1201 ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt
     1261 cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc
     1321 tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat
     1381 gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1441 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1501 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1561 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1621 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1681 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1741 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1801 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1861 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1921 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1981 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2041 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2101 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2161 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2221 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2281 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2341 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2401 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2461 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2521 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2581 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2641 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2701 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2761 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2821 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2881 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2941 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3001 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3061 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3121 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3181 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3241 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3301 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3361 gattcattaa tgcagctgga tcttcgagcg tcccaaaacc ttctcaagca aggttttcag
     3421 tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa
     3481 taacgttctt aatactaaca taactataaa aaaataaata gggacctaga cttcaggttg
     3541 tctaactcct tccttttcgg ttagagcgga tcttagctag ccgcggtacc aagcttactc
     3601 gagagatgtg attaatgaag aaatgccttt tactgactgc caaatatttt catccccaac
     3661 tcgtttttga taggaaagat tgagcacctc tggcgataaa tttttcatta ctttgtttcc
     3721 cgccccgccc gtattttgtt tctgaatctc caggttaccg ctattcagta atactttttg
     3781 gaaaattttc tgtccaccgc tatccggaag actaccaggc gcacttaaca tatgggtctg
     3841 atgtaaggaa atgatctctc gcttgatttc tgaatccatc atccctgtac gatctttgcc
     3901 gcttttagaa ttccaggcgg gtaccgcgtc aatttcatgg gctaacatgg caaggcgttg
     3961 tgcgagttta tagggttcgc cgccatcttt atgatgttgg ttatttttcc atatatcctt
     4021 aatctggcgc gctaatgtat tgacgacctc ataattatcc gggtattgcg ccagccattc
     4081 gccaacccag ccacctggtc tggcttcagg gcgtaaatca ttgcctaata actgatgtag
     4141 cgcctcggca ttatagctat ccgatgcctt aaggccaaag ccgagcttga gcgccagctc
     4201 attaacaccc acattaaatg cggcgacgtc cggttttatt tttaccgtct gtagatcgcc
     4261 atctttattg cggattttta aatgaatcat ttttcccggc tgggtcaacg attgccatgc
     4321 gcgcatttgg tcctcgacca tcgttccctc tttgccgaaa atattcgacg cggtgagtaa
     4381 cccgacggat accagtttca ggcttaccgc ctcgcccgct aaggctttgt taagcaactc
     4441 aggtttacta aaaagtgccg cagttaatac ttctttggct ttgttttcag cgccgacgtg
     4501 acgcagaagc ggatcttttt catgataggg ggaaagcacg ccatgacgta tcccgcaaaa
     4561 aagcgtttta tctttaccgt cctcatgcac actcaccgtg gacatccaca aattattggc
     4621 gtgatgaata ttcttggtat cccaactgca tacgcccttt ccctcataag cactgggaaa
     4681 gatatctttt gcgccgattt tcatctctgc tgcagggagc tgcgtgttgg tatagtgatg
     4741 cccgttatgc gtgagtgtat tttttatggt ttgccagggc tggttattaa gctgcttgac
     4801 ctgagcatct ctaaacgcta ctgccgcaaa gcgatgagct tcttttgcag gtaagccatc
     4861 ctgctgctga gtgcgtgctg caataagttc gataagattt ttcttcaatg cttttgccgc
     4921 ggcttcttct tgttttgttg ctggccggtc cgctttaact ttggctaact ccatttggtt
     4981 tgccattgac gttagaaccg ggtcttgttt acccagaacg gttgccgcga cggtaagaag
     5041 atcgcgctgt aagttataga ggttatgcag cgagtggtta gacgccttct gatgctgtag
     5101 ataattcccc catgtcgcgc cgggttctcg caggacaata atttccgggc gagcgtcggg
     5161 cgctttagcc ggcgctttgc cctggcctga gagaatctgc attccgttgt ataaggtttt
     5221 ttgtaggctt ttaaaagcct cctgggtttt tagtgaagct gagtgataga agctctgtat
     5281 ttgcattgga tccggggttt tttctccttg acgttaaagt atagaggtat attaacaatt
     5341 ttttgttgat acttttatta catttgaata agaagtaata caaaccgaaa atgttgaaag
     5401 tattagttaa agtggttatg cagtttttgc atttatatat ctgttaatag atcaaaaatc
     5461 atcgcttcgc tgattaatta ccccagaaat aaggctaaaa aactaatcgc attatcatcc
     5521 tatggttgtt aatttgattc gttcatttga aggtttgtgg ggccaggtta ctgccaattt
     5581 ttcctcttca taaccataaa agctagtatt gtagaatctt tattgttcgg agcagtgcgg
     5641 cgcgaggcac atctgcgttt caggaacgcg accggtgaag acgaggacgc acggaggaga
     5701 gtcttccttc ggagggctgt cacccgctcg gcggcttcta atccgtactt caatatagca
     5761 atgagcagtt aagcgtatta ctgaaagttc caaagagaag gtttttttag gctaagataa
     5821 tggggctctt tacatttcca caacatataa gtaagattag atatggatat gtatatggat
     5881 atgtatatgg tggtaatgcc atgtaatatg attattaaac ttctttgcgt ccatccaaaa
     5941 aaaaagtaag aatttttgaa aattcgaatt caaccctcac taaagggcgg ccgcactagt
     6001 atcgatggat tacaaggatg acgacgataa gatctgagct cttaattaac aattcttcgc
     6061 cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg tctaccttgc cagaaattta
     6121 cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt gttgacactt ctaaataagc
     6181 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
     6241 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
     6301 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctccaattc agctggcgta
     6361 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6421 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6661 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6781 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6841 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     6901 acgcatctgt gcggtatttc acaccgcata tcgaccggtc gaggagaact tctagtatat
     6961 ctacatacct aatattattg ccttattaaa aatggaatcc caacaattac atcaaaatcc
     7021 acattctctt caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta
     7081 tatttatagg ataattatac tctatttctc aacaagtaat tggttgtttg gccgagcggt
     7141 ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa ctgtgggaat actcaggtat
     7201 cgtaagatgc aagagttcga atctcttagc aaccattatt tttttcctca acataacgag
     7261 aacacacagg ggcgctatcg cacagaatca aattcgatga ctggaaattt tttgttaatt
     7321 tcagaggtcg cctgacgcat ataccttttt caactgaaaa attgggagaa aaaggaaagg
     7381 tgagagcgcc ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg
     7441 catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg
     7501 gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat
     7561 atatatttca aggatatacc attctaatgt ctgcccctaa gaagatcgtc gttttgccag
     7621 gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg
     7681 atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg
     7741 atgctacagg tgttccactt ccagatgagg cgctggaagc ctccaagaag gctgatgccg
     7801 ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag
     7861 gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg
     7921 catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact
     7981 tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag gaagacgatg
     8041 gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa
     8101 gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata
     8161 aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga
     8221 acgaattccc tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag
     8281 ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca
     8341 tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg
     8401 cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc
     8461 cagatttgcc aaagaataag gtcaacccta tcgccactat cttgtctgct gcaatgatgt
     8521 tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg
     8581 ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg
     8641 gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta aaaagattct ctttttttat
     8701 gatatttgta cataaacttt ataaatgaaa ttcataatag aaacgacacg aaattacaaa
     8761 atggaatatg ttcatagggt agacgaaact atatacgcaa tctacataca tttatcaaga
     8821 aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa ttgatactaa tggctcaacg
     8881 tgataaggaa aaagaattgc actttaacat taatattgac aaggaggagg gcaccacaca
     8941 aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc taatttatat attggaggat
     9001 tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg accatttgat
     9061 ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa
     9121 ttttactctg tcagaaacgg ccttaacgac gtagtcgata tggtgcactc tcagtacaat
     9181 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     9241 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     9301 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     9361 gata
//
