LOCUS       sequence-230355-        7532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for PmCHS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7532)
  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 7532)
  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: pJKW1548
FEATURES             Location/Qualifiers
     source          1..7532
                     /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(5264..5280)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5265..5281)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5283..5530
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5299..5317)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5548..5567)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5549..5567)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5577..5594)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5577..5593)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5586..5608)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5734..6189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5821..5840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6031..6052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6320..7123)
                     /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(7124..7339)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7363..7382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7482..7504
                     /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 aagaccgtag aggagatttg ggcggcgcag cgggcgaggg
     4081 gaccagccac ggtgctggcc atcggcactg ctgcgccggc caatgtggtg taccaggccg
     4141 attatccgga ctactacttt aggatcacca agagcgagca catgacagag ctcaaggaga
     4201 agttccgacg aatgtgtgac aagtcgatga taacgaagcg gcacatgcac ttgtcggagg
     4261 agctgttgaa aaacaaccct gacatctgtg cctacatggc cccttccctc gacgcccgcc
     4321 aagatatggt cgtggtggag gtacccaagc tcggcaagga ggcggccgcc aaggccatca
     4381 aggaatgggg ccgcccaaag tcggccatca cccacctcat cttctgcacc acctccggcg
     4441 tcgacatgcc cggcgccgat ttccagctca ccaagctact cggcctctgc ccctccgttc
     4501 gccgcaccat gctctaccag cagggctgct tcgccggcgg tacggttctc cgccttgcca
     4561 aggacctcgc cgagaacaat gcgggcgcga gggtcctcgt cgtctgctcc gagatcaccg
     4621 ccgtcacctt ccgcggcccc tcggagactc acctcgatag catggtcggc caggccctgt
     4681 tcggtgatgg tgcctctgcc atcatcgtcg gtgccgaccc cgaccccgtc atagaaaggc
     4741 cactctttca aattgtatct gcggctcaga ccatccttcc cgactcggat ggggcaatag
     4801 acggccatct ccgagaagtg ggtctaacct tccacctcct caaggacgta cctgggctca
     4861 tctcaaagaa catcgagaag agcctaaagg aggcgtttgc accgctgggc atcgacgact
     4921 ggaactcgat attttggatt gttcatccag gcgggccggc cattctagac caggtggagg
     4981 cgaagctgcg tctgaaagtg gagaagctga agacaacgag aacagttttg agtgagtacg
     5041 ggaatatgtc gagcgcttgc gtgttgttca tacttgacga gatgaggagg aacagcatgg
     5101 aagaagggaa ggcgacgacc ggtgaagggt tacattgggg agttttgttt ggttttgggc
     5161 cgggcttgac agtggagacg gtcgtcttgc atagtttgcc catcgccgag gccaactaaa
     5221 ctagtggatc ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     5281 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     5341 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     5401 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     5461 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     5521 tttaatttgc ggccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     5581 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5641 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5701 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     5761 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5821 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5881 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5941 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6001 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6061 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6121 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     6181 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     6241 agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
     6301 ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc
     6361 ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc
     6421 ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt ctatactgtt
     6481 gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt
     6541 aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt
     6601 cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc
     6661 atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct
     6721 gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc
     6781 attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag
     6841 caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct
     6901 taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa
     6961 ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg
     7021 aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac
     7081 taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc
     7141 tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     7201 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     7261 gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa
     7321 aaaaaaatga tgaattgaaa aggtggtatg gtgcactctc agtacaatct gctctgatgc
     7381 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     7441 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     7501 gaggttttca ccgtcatcac cgaaacgcgc ga
//
