LOCUS       sequence-353107-        8247 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 8247)
  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 8247)
  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..8247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(13..454)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(19..42)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    337..454
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5397..5414)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5397..5413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5406..5428)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5622..5641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5741..5763
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5801..5819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5887..5991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5992..6060,6061..6852)
                     /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(6210..6229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7023..7611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7512..7531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7765..7782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7899..7920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7935..7940,7941..7958,7959..7965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7973..7989
                     /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     7978..8000
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7997..8013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7997..8013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttatagtttt ttctccttga cgttaaagta tagaggtata ttaacaattt tttgttgata
       61 cttttatgac atttgaataa gaagtaatac aaactgaaaa tgttgaaagt attagttaaa
      121 gtggttatgc agcttttcca tttatatatc tgttaataga tcaaaaatca tcgcttcgct
      181 gattaattac cccagaaata aggctaaaaa actaatcgca ttatcatcct atggttgtta
      241 atttgattcg ttaatttgaa ggtttgtggg gccaggttac tgccaatttt tcctcttcat
      301 aaccataaaa gctagtattg tagaatcttt attgttcgga gcagtgcggc gcgaggcaca
      361 tctgcgtttc aggaacgcga ccggtgaaga cgaggacgca cggaggagag tcttccgtcg
      421 gagggctgtc gcccgctcgg cggcttctaa tccgtacttc aatatagcaa tgagcagtta
      481 agcgtattac tgaaagttcc aaagagaagg tttttttagg ctaagataat ggggctcttt
      541 acatttccac aacatataag taagattaga tatggatatg tatatggtgg taatgccatg
      601 taatatgatt attaaacttc tttgcgtcca tccaaaaaaa aagtaagaat ttttgaaaat
      661 tcaatataag gatccaaaat ggcttcagaa aaagaaatta ggagagagag attcttgaac
      721 gttttcccta aattagtaga ggaattgaac gcatcgcttt tggcttacgg tatgcctaag
      781 gaagcatgtg actggtatgc ccactcattg aactacaaca ctccaggcgg taagttaaat
      841 agaggtttgt ccgttgtgga cacgtatgct attctctcca acaagaccgt tgaacaattg
      901 gggcaagaag aatacgaaaa ggttgctatt ctaggttggt gcattgagtt gttgcaggct
      961 tacttcttgg tcgccgatga tatgatggac aagtccatta ccagaagagg ccaaccatgt
     1021 tggtacaagg ttcctgaagt tggggaaatt gccatcaatg acgcattcat gttagaggct
     1081 gctatctaca agcttttgaa atctcacttc agaaacgaaa aatactacat agatatcacc
     1141 gaattgttcc atgaagtcac cttccaaacc gaattgggcc aattgatgga cttaatcact
     1201 gcacctgaag acaaagtcga cttgagtaag ttctccctaa agaagcactc cttcatagtt
     1261 actttcaaga ctgcttacta ttctttctac ttgcctgtcg cattggctat gtacgttgcc
     1321 ggtatcacag atgaaaagga tttgaaacaa gccagagatg tcttgattcc attgggtgaa
     1381 tatttccaaa ttcaagatga ctacttagac tgcttcggta ccccagaaca gatcggtaag
     1441 atcggtacag atatccaaga taacaaatgt tcttgggtaa tcaacaaggc attagaactt
     1501 gcttccgcag aacaaagaaa gactttagac gaaaattacg gtaagaagga ctcagtcgca
     1561 gaagccaaat gcaaaaagat tttcaatgac ttgaaaatcg accagttata ccacgaatat
     1621 gaagagtctg ttgccaagga tttgaaggcc aagatctccc aagtcgacga gtctcgtggc
     1681 ttcaaagccg acgtcttaac tgcgtttttg aacaaagttt acaagagaag caaataaaga
     1741 tcttttgcca gcttactatc cttcttgaaa atatgcactc tatatctttt agttcttaat
     1801 tgcaacacat agatttgctg tataacgaat tttatgctat tttttaaatt tggagttcag
     1861 tgataaaagt gtcacagcga atttcctcac atgtagggac cgaattgttt acaagttctc
     1921 tgtaccacca tggagacatc aaaaattgaa aatctatgga aagatatgga cggtagcaac
     1981 aagaatatag cacgagccgc ggagttcatt tcgttacttt tgatatcact cacaactatt
     2041 gcgaagcgct tcagtgaaaa aatcataagg aaaagttgta aatattattg gtagtattcg
     2101 tttggtaaag tagagggggt aatttttccc ctttattttg ttcatacatt cttaaattgc
     2161 tttgcctctc cttttggaaa gctatacttc ggagcactgt tgagcgaagg ctcattagat
     2221 atattttctg tcattttcct taacccaaaa ataagggaaa gggtccaaaa agcgctcgga
     2281 caactgttga ccgtgatccg aaggactggc tatacagtgt tcacaaaata gccaagctga
     2341 aaataatgtg tagctatgtt cagttagttt ggctagcaaa gatataaaag caggtcggaa
     2401 atatttatgg gcattattat gcagagcatc aacatgataa aaaaaaacag ttgaatattc
     2461 cctcaaaaga gctcaaaatg gaattgtatg ctcaatctgt tggtgttggt gctgcatcaa
     2521 gacctctagc caattttcat caatgtgtat ggggagacaa attcattgtt tacaacccac
     2581 aatcatccca ggctggagag agagaacagg ctgaggagct taaagtagag ctaaaaagag
     2641 agcttaagga agcatcagat aattatatga ggcaactgaa aatggtagat gcaatacaac
     2701 gtttaggcat tgactattta tttgtcgaag atgttgatga agctttgaag aatttgtttg
     2761 aaatgtttga tgctttctgc aagaataatc atgacatgca cgccactgct ctatcctttc
     2821 gtcttctaag acaacatggc tatagagttt catgtgaagt ttttgaaaag tttaaggatg
     2881 gcaaagatgg atttaaggtt ccaaatgagg atggagccgt tgcagtcctt gaattcttcg
     2941 aagccactca tttgagagtc catggagaag acgtccttga taatgctttt gtcttcacta
     3001 ggaactactt ggaatcagtc tatgcaactt tgaacgatcc aaccgcgaat caagtccaca
     3061 acgcattgaa tgagttctct tttaggagag gattgccacg tgtggaagca aggaagtaca
     3121 tatcaatcta cgagcaatac gcatctcatc acaaaggctt gttaaaactt gctaagctgg
     3181 atttcaactt ggtacaagct ttgcacagaa gggagctgag tgaagattct aggtggtgga
     3241 agactttaca agtgcccaca gagctatcat tcgttagaga taggttggtg gagtcctact
     3301 tttgggcttc gggatcttat ttcgaaccga attattcggt agctaggatg attttagcaa
     3361 aaggattagc tgtattatct cttatggacg atgtgtatga tgcatatggt acttttgagg
     3421 aattacaagt gttcacagat gcaatcgaaa ggtgggatgc ttcatgttta gataaacttc
     3481 cagagtacat gaaaatagta tacaaggccc ttttggatgt gtttgaggaa gttgacgagg
     3541 aggtgatcaa gctaggtgca ccatataggg tctactatgg aaaagaagcc atgaaatatg
     3601 ccgctagagc ttacatggaa gaggcccaat ggagggagca aaagcacaaa cccacaacta
     3661 aggagtatat gaagctggca acaaagacct gtggctatat aactctaata atattatcat
     3721 ttcttggagt ggaagagggc attgtgacta aagaagcctt cgattgggtt ttctccagac
     3781 ctcctttcgt cgaggctact ttaatcattg ctaggttgat caatgatatt acaggatgtg
     3841 aatttgagaa taaaagggag cacgttagaa ctgcagtcga atgctacatg gaagagcaca
     3901 aagtgggaaa gcaagaggtg gtctctgaat tttacaacca aatggagtca gcatggaagg
     3961 acataaatga gtgcttgcta agaccagctg aatttccaat ccctctactt aatcttattt
     4021 taaattctgt cagaacactt gaggttattt acaaagaggg cgattcctat acacacgtgg
     4081 gtcctgcaat gcaaaacatc atcaagcagt tgtacctaca ccctgttcca tattaactcg
     4141 agatagatta atttaaacag tatatgtaca gttttatata tatatatata tatatataca
     4201 tatataaaga aacctgtgcg ttttttgtat tttcaaatac atttagtttt ggcgccactt
     4261 ctataaaagg tctcatgata ttgttactct agagatccca atacaacaga tcacgtgatc
     4321 ttttgtaaga tgaagttgaa gtgagtgttg caccgtgcca atgcaggtgg ctattagatt
     4381 aaatatgtga tttgttctat taagtttcct gtataattaa tggggagcgc tgattctctt
     4441 ttggtacgct tcccatccag catttctgta tctttcacct tcaaccttag gatctctacc
     4501 cttggcgaaa agtcctctgc caacaatgat gatatctgat ccaccactta caacttcgtc
     4561 gacggttctg tactgctgac ccaatgcatc gcctttgtcg tctaaaccta cacctggggt
     4621 catgattagc caatcaaacc cttcttctct tcctcccata tcgttctgag caatgaaccc
     4681 aataacgaaa tctttatcac tctttgcaat atcaacggta cccttagtat attcaccgtg
     4741 tgctagagaa cccttggaag acaattcagc aagcatcaat aatccccttg gttctttggt
     4801 gacctcttgc gcaccttgtt tcaagccagc aacaatacca gcaccagtaa ccccgtgggc
     4861 gttggtgata tcagaccatt ctgcgatacg gtaaacgccc gatgtatatt gtaatttgac
     4921 tgtgttaccg atatcggcga attttctgtc ctcaaatatc aagaacttgt atttctctgc
     4981 caatgctttc aatggaacga cagtaccctc ataactgaaa tcatccaaga tatcaacgtg
     5041 tgttttcaaa aggcaaatgt atggacccaa cgtttcaaca agtttcaata gctcatcagt
     5101 cgaacgaacg tcaagagaag cacacaaatt ggtcttcttt tcatccatta aacgtaaaag
     5161 tttcgatgca accggacttg catgagtctc agctctactg gtatatgatt ttgtggacat
     5221 gttttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca
     5281 atttcatgtt tcttcaacac cacatatgcg tatatatacc aatctaagtc tgtgctcctt
     5341 ccttcgttct tccttctgct cggagattac cgaatcaaaa aaatttaaat gaattcactg
     5401 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5461 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5521 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     5581 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     5641 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5701 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5761 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7141 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7681 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7741 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7801 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7861 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7921 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7981 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcatt
     8041 taaatcatta cgaccgagat tcccgggtaa taactgatat aattaaattg aagctctaat
     8101 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca
     8161 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc
     8221 ccttcccttt gcaaatagtc ctactag
//
