LOCUS       sequence-230356-        7541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for PmSPS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7541)
  AUTHORS   Pluskal T, Torrens-Spence MP, Fallon TR, De Abreu A, Shi CH, Weng JK
  TITLE     The biosynthetic origin of psychoactive kavalactones in kava.
  JOURNAL   Nat Plants. 2019 Jul 22. pii: 10.1038/s41477-019-0474-0. doi: 
            10.1038/s41477-019-0474-0.
  PUBMED    31332312
REFERENCE   2  (bases 1 to 7541)
  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 Plants.
            2019 Jul 22. pii: 10.1038/s41477-019-0474-0. doi: 
            10.1038/s41477-019-0474-0."
COMMENT     Sequence Label: pJKW1538
FEATURES             Location/Qualifiers
     source          1..7541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      69..1411
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    401..448
                     /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)."
     promoter        1438..1542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1543..1611,1612..2403)
                     /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(1761..1780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2574..3162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3063..3082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3316..3333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3450..3471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3486..3491,3492..3509,3510..3516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3540
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3529..3551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3548..3564
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3583..3603
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3585..3603
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3621..4019
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(5273..5289)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5274..5290)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5292..5539
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5308..5326)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5557..5576)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5558..5576)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5586..5603)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5586..5602)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5595..5617)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5743..6198
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5830..5849)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6040..6061
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6329..7132)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7133..7348)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7372..7391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7491..7513
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     3481 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3601 agggaacaaa agctggagct catagcttca aaatgtttct actccttttt tactcttcca
     3661 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
     3721 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
     3781 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
     3841 acgtttcttt ttcttgaaaa tttttttttt gatttttttc tctttcgatg acctcccatt
     3901 gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc
     3961 tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagt
     4021 tttctagaac tagtatgtcg aagacggtgg aggatcgggc agcgcagcgg gcaaaggggc
     4081 cggcaacagt gctggccatc ggcacggcta cgccggccaa tgtggtgtac cagaccgatt
     4141 atccggacta ctacttcagg gtcaccaaga gcgagcatat gaccaaactc aagaacaagt
     4201 ttcaacgcat gtgcgacagg tcgacgataa agaagaggta catggttttg acagaggagc
     4261 tgctagagaa gaatctgagt ttgtgcacct acatggaacc ctccctcgac gcccgccaag
     4321 acattctcgt gccggaggtc cccaagctcg gcaaggaggc cgccgacgag gccatcgccg
     4381 aatggggacg ccccaagtcg gaaatcaccc acctcatctt ttgcactacc tgcggcgtcg
     4441 acatgcccgg cgccgactac cagctcacca agctcctcgg tctccgctcc tccgtccgtc
     4501 gcaccatgct ctatcagcag ggatgctttg gcggaggcac cgttctccgc ctcgccaagg
     4561 acctcgccga gaacaacgct ggtgcccgcg tcctcgtcgt ctgctccgag atcaccactg
     4621 ccgtcaactt ccgagggcct tccgacaccc acctcgactt attggtcggc ttagccctgt
     4681 tcggcgacgg tgcggccgcg gtcatagtcg gtgcggatcc agatcctacc ctcgagcggc
     4741 cgctctttca aatcgtatct ggagcacaga cgattctacc ggactcggag ggggccatca
     4801 acggccatct ccgggaggtg gggctaacca tccgcctact caaggacgta cctgggcttg
     4861 tgtcgatgaa cattgagaag tgcctcatgg aggcgtttgc accgatgggc atccacgact
     4921 ggaactccat cttttggata gcccatcccg gggggcccac catactagac caagtggagg
     4981 ccaagctggg tctaaaggag gagaagctca agtcgacgag ggctgttctg agggagtatg
     5041 gcaacatgtc tagcgcctgt gtcttgttca tactggacga ggtaaggaag aggagcatgg
     5101 aggaggggaa gacgacaacc ggtgaggggt tcgattgggg agttctattc ggctttgggc
     5161 ctggcttcac agtggagacc gtcgtcttgc acagcatgcc catccccaaa gccgatgaag
     5221 gcagataaac tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg
     5281 tcgacctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     5341 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     5401 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     5461 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     5521 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     5581 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5641 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5701 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     5761 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5821 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5881 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5941 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6001 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6061 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6121 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6181 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6241 acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     6301 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     6361 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     6421 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     6481 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     6541 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     6601 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     6661 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     6721 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     6781 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     6841 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     6901 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     6961 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     7021 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     7081 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     7141 ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     7201 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     7261 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     7321 aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca gtacaatctg
     7381 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     7441 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7501 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg a
//
