LOCUS       sequence-231689-        6048 bp    DNA     circular SYN 15-APR-2025
DEFINITION  E.coli expression plasmid for Pm4CL1 and PmCHS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  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 6048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJKW1659
FEATURES             Location/Qualifiers
     source          1..6048
                     /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             1790..1812
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             3490..3512
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3827..3843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3827..3843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3840..3862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3851..3867
                     /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(3875..3881,3882..3899,3900..3905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3920..3941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4058..4075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4229..4817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4309..4328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4988..5779,5780..5848))
                     /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     5611..5630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5849..5953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6021..6039
                     /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 agctcattag
      601 ttggcctcgg cgatgggcaa actatgcaag acgaccgtct ccactgtcaa gcccggccca
      661 aaaccaaaca aaactcccca atgtaaccct tcaccggtcg tcgccttccc ttcttccatg
      721 ctgttcctcc tcatctcgtc aagtatgaac aacacgcaag cgctcgacat attcccgtac
      781 tcactcaaaa ctgttctcgt tgtcttcagc ttctccactt tcagacgcag cttcgcctcc
      841 acctggtcta gaatggccgg cccgcctgga tgaacaatcc aaaatatcga gttccagtcg
      901 tcgatgccca gcggtgcaaa cgcctccttt aggctcttct cgatgttctt tgagatgagc
      961 ccaggtacgt ccttgaggag gtggaaggtt agacccactt ctcggagatg gccgtctatt
     1021 gccccatccg agtcgggaag gatggtctga gccgcagata caatttgaaa gagtggcctt
     1081 tctatgacgg ggtcggggtc ggcaccgacg atgatggcag aggcaccatc accgaacagg
     1141 gcctggccga ccatgctatc gaggtgagtc tccgaggggc cgcggaaggt gacggcggtg
     1201 atctcggagc agacgacgag gaccctcgcg cccgcattgt tctcggcgag gtccttggca
     1261 aggcggagaa ccgtaccgcc ggcgaagcag ccctgctggt agagcatggt gcggcgaacg
     1321 gaggggcaga ggccgagtag cttggtgagc tggaaatcgg cgccgggcat gtcgacgccg
     1381 gaggtggtgc agaagatgag gtgggtgatg gccgactttg ggcggcccca ttccttgatg
     1441 gccttggcgg ccgcctcctt gccgagcttg ggtacctcca ccacgaccat atcttggcgg
     1501 gcgtcgaggg aaggggccat gtaggcacag atgtcagggt tgtttttcaa cagctcctcc
     1561 gacaagtgca tgtgccgctt cgttatcatc gacttgtcac acattcgtcg gaacttctcc
     1621 ttgagctctg tcatgtgctc gctcttggtg atcctaaagt agtagtccgg ataatcggcc
     1681 tggtacacca cattggccgg cgcagcagtg ccgatggcca gcaccgtggc tggtcccctc
     1741 gcccgctgcg ccgcccaaat ctcctctacg gtcttcgaca ttatggcagt ctccttctta
     1801 aagttaaaca aacaaccgta ccggatcccc tgcagggcta cttgggtata ccagctgcca
     1861 atcttcccct caaatccttc ctcaaaatct tcccagaggg tgctttagga attgaatcaa
     1921 cgaaaaagac tcgattgatt cgcttgtaga acaccacctg cttgctaatg aactgcttga
     1981 tctcatcctc actgatgact gatccattgg aggtcaccac aaatgccact ggcacttccc
     2041 ctgctgcctc atctttcatc gggacaacag cggcatcagc aatgttaggg tgagtaatga
     2101 gaatggcttc gagctcggca ggggcgactt ggaagccctt gtacttaatg atctccttca
     2161 acctatcgac aatgaagagc tcctcgtcgt cgtcgacata gccaatgtcg ccggtgtgca
     2221 gccacccttc cttgtcgatc gtcctctgcg tagccgccgc gtcattaaga taccctttca
     2281 tgatttggga gcctcggatg caaatttcgc cgggttggtt tctgggcagg taggcaccgg
     2341 tttctgggtc gacgatcttg agctgggcgt tcctgaccac ggtgccgcaa gaaccagact
     2401 tgatctcgaa gggctccttg gcgaaggcca aacacatgga cagcactggc cctgcctccg
     2461 tcatcccata gccctggccg agcttggcgt tgggaagctt ggcccggacg gcgtcttcga
     2521 gctccttccc catcggggcg gcgccggaca tcaccgtccg aatggacgac aagtcgtact
     2581 tgtcgacgag cgggctcttg gcgatggcaa gaacgatggg cggaacgatg ggcgcaatgg
     2641 tcaccttgta cttctgcacc agctccatca gcgccgtgat ctcgaacttc tgcatgatga
     2701 ggatcgccgc cccgacgcgc aggccgcaga agagcaccga gttgagggag tagatgtgga
     2761 agaggggaag aacgcagagc aggacgtcgt ctggccggaa gtatagattc gggttctcgc
     2821 cgtcgacttg ctgggcaacg ctcgtcacct ggctgcggtg tgtgagcatc actcctttag
     2881 ggaggccggt ggtgccggag gagtatggca gcgccaccac gtcgtcgggg tcaatatcca
     2941 cctcgggcag ctccgactcg tcggcgccgg acagctcgga gaagtgggag caccccgggg
     3001 gcggcgcgtc gacgcacacc actttgacgc cgcgctcctc ggccaggtcc ctgaccttgt
     3061 cgacgtaggc ggacatggtg acgatgagct tcgcgccgga ggccttgacc tgcttggcga
     3121 tctcctgcgg cgtgtagaag gggttggcgg tggtggacat ggccccgcgg aaggaggcgc
     3181 cgaggaaggc gaagacgaac tccggcgagt tggggaggag gatcatgatc acctcccgct
     3241 gctgcacccc catccggtgc agcccggcgg ccaccctcct cgccgtcgtc tccacctcgc
     3301 cgtacgtgta cacctcgccg gtgctgccat tgatgagaca aggattgtcc cggtaagcac
     3361 caatgttctg gaaacaataa ttgtgcaacg acatgtcgtt cttgatctca atgtccggca
     3421 gcttagaccg gtatacacat cgatcagtag caatagtgtc tactaccatc ttcatcctcg
     3481 agatgcagtt ctccttctta aagttaaaca aacggcatat tccaccagct atttgttagt
     3541 gaataaaagg ttgcctgtaa aattacaaaa accttacgca gcgtcaagcg gaatcgtgtc
     3601 aatcattgcg acaaatcaat cctgtgccta agcattacgc gactgactcg cctcactctt
     3661 cctttgggct tgagaccaca tggggtcggc gccccgaatt ttaaagggca attacgataa
     3721 aaatgtgatt ttcatcacga tttaggtgca gccatttcgt cgagatcaag tttaggacaa
     3781 aattttgcca gttggtgtta atgttttgtt agaatccgta atcatggtca tagctgtttc
     3841 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3901 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4021 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4081 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4141 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6001 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
