LOCUS       sequence-230380-        6057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli expression plasmid for Pm4CL1 and PmSPS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6057)
  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 6057)
  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: pJKW1574
FEATURES             Location/Qualifiers
     source          1..6057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             543..565
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1799..1821
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             3499..3521
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3836..3852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3836..3852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3849..3871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3860..3876
                     /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)."
     promoter        complement(join(3884..3890,3891..3908,3909..3914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3929..3950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4067..4084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4238..4826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4318..4337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4997..5788,5789..5857))
                     /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     5620..5639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5858..5962)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6030..6048
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctatgagaa gagaaaagaa
      421 aaccgccgat cctgtccacc gcattactgc aaggtagtgg acaagaccgg cggtcttaag
      481 ttttttggct gaatgattcg agtggatcca agcttgcggc cgcgtcgaca ccggtatgta
      541 ggtctccttc ttaaagttaa acaaatcagg cgatcaagct tttaattaag agctcattat
      601 ctgccttcat cggctttggg gatgggcatg ctgtgcaaga cgacggtctc cactgtgaag
      661 ccaggcccaa agccgaatag aactccccaa tcgaacccct caccggttgt cgtcttcccc
      721 tcctccatgc tcctcttcct tacctcgtcc agtatgaaca agacacaggc gctagacatg
      781 ttgccatact ccctcagaac agccctcgtc gacttgagct tctcctcctt tagacccagc
      841 ttggcctcca cttggtctag tatggtgggc cccccgggat gggctatcca aaagatggag
      901 ttccagtcgt ggatgcccat cggtgcaaac gcctccatga ggcacttctc aatgttcatc
      961 gacacaagcc caggtacgtc cttgagtagg cggatggtta gccccacctc ccggagatgg
     1021 ccgttgatgg ccccctccga gtccggtaga atcgtctgtg ctccagatac gatttgaaag
     1081 agcggccgct cgagggtagg atctggatcc gcaccgacta tgaccgcggc cgcaccgtcg
     1141 ccgaacaggg ctaagccgac caataagtcg aggtgggtgt cggaaggccc tcggaagttg
     1201 acggcagtgg tgatctcgga gcagacgacg aggacgcggg caccagcgtt gttctcggcg
     1261 aggtccttgg cgaggcggag aacggtgcct ccgccaaagc atccctgctg atagagcatg
     1321 gtgcgacgga cggaggagcg gagaccgagg agcttggtga gctggtagtc ggcgccgggc
     1381 atgtcgacgc cgcaggtagt gcaaaagatg aggtgggtga tttccgactt ggggcgtccc
     1441 cattcggcga tggcctcgtc ggcggcctcc ttgccgagct tggggacctc cggcacgaga
     1501 atgtcttggc gggcgtcgag ggagggttcc atgtaggtgc acaaactcag attcttctct
     1561 agcagctcct ctgtcaaaac catgtacctc ttctttatcg tcgacctgtc gcacatgcgt
     1621 tgaaacttgt tcttgagttt ggtcatatgc tcgctcttgg tgaccctgaa gtagtagtcc
     1681 ggataatcgg tctggtacac cacattggcc ggcgtagccg tgccgatggc cagcactgtt
     1741 gccggcccct ttgcccgctg cgctgcccga tcctccaccg tcttcgacat tatggcagtc
     1801 tccttcttaa agttaaacaa acaaccgtac cggatcccct gcagggctac ttgggtatac
     1861 cagctgccaa tcttcccctc aaatccttcc tcaaaatctt cccagagggt gctttaggaa
     1921 ttgaatcaac gaaaaagact cgattgattc gcttgtagaa caccacctgc ttgctaatga
     1981 actgcttgat ctcatcctca ctgatgactg atccattgga ggtcaccaca aatgccactg
     2041 gcacttcccc tgctgcctca tctttcatcg ggacaacagc ggcatcagca atgttagggt
     2101 gagtaatgag aatggcttcg agctcggcag gggcgacttg gaagcccttg tacttaatga
     2161 tctccttcaa cctatcgaca atgaagagct cctcgtcgtc gtcgacatag ccaatgtcgc
     2221 cggtgtgcag ccacccttcc ttgtcgatcg tcctctgcgt agccgccgcg tcattaagat
     2281 accctttcat gatttgggag cctcggatgc aaatttcgcc gggttggttt ctgggcaggt
     2341 aggcaccggt ttctgggtcg acgatcttga gctgggcgtt cctgaccacg gtgccgcaag
     2401 aaccagactt gatctcgaag ggctccttgg cgaaggccaa acacatggac agcactggcc
     2461 ctgcctccgt catcccatag ccctggccga gcttggcgtt gggaagcttg gcccggacgg
     2521 cgtcttcgag ctccttcccc atcggggcgg cgccggacat caccgtccga atggacgaca
     2581 agtcgtactt gtcgacgagc gggctcttgg cgatggcaag aacgatgggc ggaacgatgg
     2641 gcgcaatggt caccttgtac ttctgcacca gctccatcag cgccgtgatc tcgaacttct
     2701 gcatgatgag gatcgccgcc ccgacgcgca ggccgcagaa gagcaccgag ttgagggagt
     2761 agatgtggaa gaggggaaga acgcagagca ggacgtcgtc tggccggaag tatagattcg
     2821 ggttctcgcc gtcgacttgc tgggcaacgc tcgtcacctg gctgcggtgt gtgagcatca
     2881 ctcctttagg gaggccggtg gtgccggagg agtatggcag cgccaccacg tcgtcggggt
     2941 caatatccac ctcgggcagc tccgactcgt cggcgccgga cagctcggag aagtgggagc
     3001 accccggggg cggcgcgtcg acgcacacca ctttgacgcc gcgctcctcg gccaggtccc
     3061 tgaccttgtc gacgtaggcg gacatggtga cgatgagctt cgcgccggag gccttgacct
     3121 gcttggcgat ctcctgcggc gtgtagaagg ggttggcggt ggtggacatg gccccgcgga
     3181 aggaggcgcc gaggaaggcg aagacgaact ccggcgagtt ggggaggagg atcatgatca
     3241 cctcccgctg ctgcaccccc atccggtgca gcccggcggc caccctcctc gccgtcgtct
     3301 ccacctcgcc gtacgtgtac acctcgccgg tgctgccatt gatgagacaa ggattgtccc
     3361 ggtaagcacc aatgttctgg aaacaataat tgtgcaacga catgtcgttc ttgatctcaa
     3421 tgtccggcag cttagaccgg tatacacatc gatcagtagc aatagtgtct actaccatct
     3481 tcatcctcga gatgcagttc tccttcttaa agttaaacaa acggcatatt ccaccagcta
     3541 tttgttagtg aataaaaggt tgcctgtaaa attacaaaaa ccttacgcag cgtcaagcgg
     3601 aatcgtgtca atcattgcga caaatcaatc ctgtgcctaa gcattacgcg actgactcgc
     3661 ctcactcttc ctttgggctt gagaccacat ggggtcggcg ccccgaattt taaagggcaa
     3721 ttacgataaa aatgtgattt tcatcacgat ttaggtgcag ccatttcgtc gagatcaagt
     3781 ttaggacaaa attttgccag ttggtgttaa tgttttgtta gaatccgtaa tcatggtcat
     3841 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3901 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3961 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4021 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4081 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4141 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5281 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5581 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5881 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5941 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6001 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
