LOCUS       sequence-360444-        8246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing ERG20 (GAL10 promoter) and 
            patchoulol synthase from Pogostemon cablin (PcPTS; GAL7 promoter). 
            Contains uracil auxotrophic marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  AUTHORS   Cheah LC, Liu L, Stark T, Plan MR, Peng B, Lu Z, Schenk G, Sainsbury
            F, Vickers CE
  TITLE     Metabolic flux enhancement from the translational fusion of terpene 
            synthases is linked to terpene synthase accumulation.
  JOURNAL   Metab Eng. 2023 May;77:143-151. doi: 10.1016/j.ymben.2023.03.012. 
            Epub 2023 Mar 28.
  PUBMED    36990382
REFERENCE   2  (bases 1 to 8246)
  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: "Metab 
            Eng."; date: "2023-05"; volume: "77:143-151. doi"; pages: " 
            10.1016/j.ymben.2023.03.012. Epub 2023 Mar 28"
COMMENT     Sequence Label: Coexpressed Free PcPTS
FEATURES             Location/Qualifiers
     source          1..8246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(102..543)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(108..131)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    426..543
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5485..5502)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5485..5501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5494..5516)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5710..5729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5829..5851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5889..5907)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5975..6079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6080..6148,6149..6940)
                     /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(6298..6317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7111..7699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7600..7619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7853..7870
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7987..8008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8023..8028,8029..8046,8047..8053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8061..8077
                     /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     8066..8088
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8085..8101
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8085..8101
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
       61 tcccttccct ttgcaaatag tcctactagt tatagttttt tctccttgac gttaaagtat
      121 agaggtatat taacaatttt ttgttgatac ttttatgaca tttgaataag aagtaataca
      181 aaccgaaaat gttgaaagta ttagttaaag tggttatgca gcttttgcat ttatatatct
      241 gttaatagat caaaaatcat cgcttcgctg attaattacc ccagaaataa ggctaaaaaa
      301 ctaatcgcat tattatccta tggttgttaa tttgattcgt tgatttgaag gtttgtgggg
      361 ccaggttact gccaattttt cctcttcata accataaaag ctagtattgt agaatcttta
      421 ttgttcggag cagtgcggcg cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac
      481 caggacgcac ggaggagagt cttccgtcgg agggctgtcg cccgctcggc ggcttctaat
      541 ccgtacttca atatagcaat gagcagttaa gcgtattact gaaagttcca aagagaaggt
      601 ttttttaggc taagataatg gggctcttta catttccaca acatataagt aagattagat
      661 atggatatgt atatggtggt attgccatgt aatatgatta ttaaacttct ttgcgtccat
      721 ccaaaaaaaa aagtaagaat ttttgaaaat tcaatataag gatccaaaat ggcttcagaa
      781 aaagaaatta ggagagagag attcttgaac gttttcccta aattagtaga ggaattgaac
      841 gcatcgcttt tggcttacgg tatgcctaag gaagcatgtg actggtatgc ccactcattg
      901 aactacaaca ctccaggcgg taagttaaat agaggtttgt ccgttgtgga cacgtatgct
      961 attctctcca acaagaccgt tgaacaattg gggcaagaag aatacgaaaa ggttgctatt
     1021 ctaggttggt gcattgagtt gttgcaggct tacttcttgg tcgccgatga tatgatggac
     1081 aagtccatta ccagaagagg ccaaccatgt tggtacaagg ttcctgaagt tggggaaatt
     1141 gccatcaatg acgcattcat gttagaggct gctatctaca agcttttgaa atctcacttc
     1201 agaaacgaaa aatactacat agatatcacc gaattgttcc atgaagtcac cttccaaacc
     1261 gaattgggcc aattgatgga cttaatcact gcacctgaag acaaagtcga cttgagtaag
     1321 ttctccctaa agaagcactc cttcatagtt actttcaaga ctgcttacta ttctttctac
     1381 ttgcctgtcg cattggctat gtacgttgcc ggtatcacag atgaaaagga tttgaaacaa
     1441 gccagagatg tcttgattcc attgggtgaa tatttccaaa ttcaagatga ctacttagac
     1501 tgcttcggta ccccagaaca gatcggtaag atcggtacag atatccaaga taacaaatgt
     1561 tcttgggtaa tcaacaaggc attagaactt gcttccgcag aacaaagaaa gactttagac
     1621 gaaaattacg gtaagaagga ctcagtcgca gaagccaaat gcaaaaagat tttcaatgac
     1681 ttgaaaatcg accagttata ccacgaatat gaagagtctg ttgccaagga tttgaaggcc
     1741 aagatctccc aagtcgacga gtctcgtggc ttcaaagccg acgtcttaac tgcgtttttg
     1801 aacaaagttt acaagagaag caaataaaga tcttttgcca gcttactatc cttcttgaaa
     1861 atatgcactc tatatctttt agttcttaat tgcaacacat agatttgctg tataacgaat
     1921 tttatgctat tttttaaatt tggagttcag tgataaaagt gtcacagcga atttcctcac
     1981 atgtagggac cgaattgttt acaagttctc tgtaccacca tggagacatc aaaaattgaa
     2041 aatctatgga aagatatgga cggtagcaac aagaatatag cacgagccgc ggagttcatt
     2101 tcgttacttt tgatatcact cacaactatt gcgaagcgct tcagtgaaaa aatcataagg
     2161 aaaagttgta aatattattg gtagtattcg tttggtaaag tagagggggt aatttttccc
     2221 ctttattttg ttcatacatt cttaaattgc tttgcctctc cttttggaaa gctatacttc
     2281 ggagcactgt tgagcgaagg ctcattagat atattttctg tcattttcct taacccaaaa
     2341 ataagggaaa gggtccaaaa agcgctcgga caactgttga ccgtgatccg aaggactggc
     2401 tatacagtgt tcacaaaata gccaagctga aaataatgtg tagctatgtt cagttagttt
     2461 ggctagcaaa gatataaaag caggtcggaa atatttatgg gcattattat gcagagcatc
     2521 aacatgataa aaaaaaacag ttgaatattc cctcaaaaga gctcaaaatg gaattgtatg
     2581 ctcaatctgt tggtgttggt gctgcatcaa gacctctagc caattttcat caatgtgtat
     2641 ggggagacaa attcattgtt tacaacccac aatcatccca ggctggagag agagaacagg
     2701 ctgaggagct taaagtagag ctaaaaagag agcttaagga agcatcagat aattatatga
     2761 ggcaactgaa aatggtagat gcaatacaac gtttaggcat tgactattta tttgtcgaag
     2821 atgttgatga agctttgaag aatttgtttg aaatgtttga tgctttctgc aagaataatc
     2881 atgacatgca cgccactgct ctatcctttc gtcttctaag acaacatggc tatagagttt
     2941 catgtgaagt ttttgaaaag tttaaggatg gcaaagatgg atttaaggtt ccaaatgagg
     3001 atggagccgt tgcagtcctt gaattcttcg aagccactca tttgagagtc catggagaag
     3061 acgtccttga taatgctttt gtcttcacta ggaactactt ggaatcagtc tatgcaactt
     3121 tgaacgatcc aaccgcgaat caagtccaca acgcattgaa tgagttctct tttaggagag
     3181 gattgccacg tgtggaagca aggaagtaca tatcaatcta cgagcaatac gcatctcatc
     3241 acaaaggctt gttaaaactt gctaagctgg atttcaactt ggtacaagct ttgcacagaa
     3301 gggagctgag tgaagattct aggtggtgga agactttaca agtgcccaca gagctatcat
     3361 tcgttagaga taggttggtg gagtcctact tttgggcttc gggatcttat ttcgaaccga
     3421 attattcggt agctaggatg attttagcaa aaggattagc tgtattatct cttatggacg
     3481 atgtgtatga tgcatatggt acttttgagg aattacaagt gttcacagat gcaatcgaaa
     3541 ggtgggatgc ttcatgttta gataaacttc cagagtacat gaaaatagta tacaaggccc
     3601 ttttggatgt gtttgaggaa gttgacgagg aggtgatcaa gctaggtgca ccatataggg
     3661 tctactatgg aaaagaagcc atgaaatatg ccgctagagc ttacatggaa gaggcccaat
     3721 ggagggagca aaagcacaaa cccacaacta aggagtatat gaagctggca acaaagacct
     3781 gtggctatat aactctaata atattatcat ttcttggagt ggaagagggc attgtgacta
     3841 aagaagcctt cgattgggtt ttctccagac ctcctttcgt cgaggctact ttaatcattg
     3901 ctaggttgat caatgatatt acaggatgtg aatttgagaa taaaagggag cacgttagaa
     3961 ctgcagtcga atgctacatg gaagagcaca aagtgggaaa gcaagaggtg gtctctgaat
     4021 tttacaacca aatggagtca gcatggaagg acataaatga gtgcttgcta agaccagctg
     4081 aatttccaat ccctctactt aatcttattt taaattctgt cagaacactt gaggttattt
     4141 acaaagaggg cgattcctat acacacgtgg gtcctgcaat gcaaaacatc atcaagcagt
     4201 tgtacctaca ccctgttcca tattaactcg agatagatta atttaaacag tatatgtaca
     4261 gttttatata tatatatata tatatacata tataaagaaa cctgtgcgtt ttttgtattt
     4321 tcaaatacat ttagttttgg cgccacttct ataaaaggtc tcatgatatt gttactctag
     4381 agatcccaat acaacagatc acgtgatctt tttgtaagat gaagttgaag tagtgttgca
     4441 ccgtgccaat gcaggtggct attagattaa atatgtgatt tgttctatta agtttcctgt
     4501 ataattaatg gggagcgctg attctctttt ggtacgcttc ccatccagca tttctgtatc
     4561 tttcaccttc aaccttagga tctctaccct tggcgaaaag tcctctgcca acaatgatga
     4621 tatctgatcc accacttaca acttcgtcga cggttctgta ctgctgaccc aatgcatcgc
     4681 ctttgtcgtc taaacctaca cctggggtca tgattagcca atcaaaccct tcttctcttc
     4741 ctcccatatc gttctgagca atgaacccaa taacgaaatc tttatcactc tttgcaatat
     4801 caacggtacc cttagtatat tcaccgtgtg ctagagaacc cttggaagac aattcagcaa
     4861 gcatcaataa tccccttggt tctttggtga cctcttgcgc accttgtttc aagccagcaa
     4921 caataccagc accagtaacc ccgtgggcgt tggtgatatc agaccattct gcgatacggt
     4981 aaacgcccga tgtatattgt aatttgactg tgttaccgat atcggcgaat tttctgtcct
     5041 caaatatcaa gaacttgtat ttctctgcca atgctttcaa tggaacgaca gtaccctcat
     5101 aactgaaatc atccaagata tcaacgtgtg ttttcaaaag gcaaatgtat ggacccaacg
     5161 tttcaacaag tttcaatagc tcatcagtcg aacgaacgtc aagagaagca cacaaattgg
     5221 tcttcttttc atccattaaa cgtaaaagtt tcgatgcaac cggacttgca tgagtctcag
     5281 ctctactggt atatgatttt gtggacatgt tttatcttcg tttcctgcag gtttttgttc
     5341 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     5401 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     5461 aatcaaaaaa atttaaatga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5521 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5581 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5641 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5701 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5761 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5821 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5881 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5941 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6001 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6061 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6121 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6181 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6241 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6301 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6361 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6421 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6481 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6541 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6601 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6661 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6721 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6781 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6841 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6901 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6961 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7021 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7081 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7141 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7201 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7261 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7321 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7381 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7441 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7501 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7561 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7621 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7681 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7741 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7801 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7861 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7921 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7981 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     8041 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     8101 ccatgattac gccaagcttg catgcattta aatcattacg accgagattc ccgggtaata
     8161 actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata
     8221 atacagtttt ttagttttgc tggccg
//
