LOCUS       sequence-353111-        7555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative plasmid for expressing fusion protein 
            ERG20(F96W-N127W)-ClLIS1, with linker sequence AG4TGGA (GAL10 
            promoter). Contains uracil auxotrophic marker (K. lactis URA3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  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 7555)
  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: Linked Free ClLIS1 (M)
FEATURES             Location/Qualifiers
     source          1..7555
                     /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(4705..4722)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4705..4721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4714..4736)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4930..4949)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5049..5071
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5109..5127)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5195..5299
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5300..5368,5369..6160)
                     /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(5518..5537)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6331..6919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6820..6839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7073..7090
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7207..7228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7243..7248,7249..7266,7267..7273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7281..7297
                     /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     7286..7308
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7305..7321
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7305..7321
                     /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 taagctaaat
      841 agaggtttgt ccgttgtgga cacgtatgct attctctcca acaagaccgt tgaacaattg
      901 gggcaagaag aatacgaaaa ggttgccatt ctaggttggt gcattgagtt gttgcaggct
      961 tactggttgg tcgccgatga tatgatggac aagtccatta ccagaagagg ccaaccatgt
     1021 tggtacaagg ttcctgaagt tggggaaatt gccatctggg acgcattcat gttagaggct
     1081 gctatctaca agcttttgaa atctcacttc agaaacgaaa aatactacat agatatcacc
     1141 gaattgttcc atgaggtcac cttccaaacc gaattgggcc aattgatgga cttaatcact
     1201 gcacctgaag acaaagtcga cttgagtaag ttctccctaa agaagcactc cttcatagtt
     1261 actttcaaga ctgcttacta ttctttctac ttgcctgtcg cattggccat gtacgttgcc
     1321 ggtatcacgg atgaaaagga tttgaaacaa gccagagatg tcttgattcc attgggtgaa
     1381 tacttccaaa ttcaagatga ctacttagac tgcttcggta ccccagaaca gatcggtaag
     1441 atcggtacag atatccaaga taacaaatgt tcttgggtaa tcaacaaggc attagaactt
     1501 gcttccgcag aacaaagaaa gactttagac gaaaattacg gtaagaagga ctcagtcgca
     1561 gaagccaaat gcaaaaagat tttcaatgac ttgaaaattg aacagctata ccacgaatat
     1621 gaagagtcta ttgccaagga tttgaaggcc aaaatttctc aggtcgatga gtctcgtggc
     1681 ttcaaagctg atgtcttaac tgcgttcttg aacaaagttt acaagagaag caaaagatct
     1741 gctggtggag gaggtactgg tggtgctgag ctcatgagaa gatcagctaa ctatcaacca
     1801 tccatttggg accacgactt tttacaatcc ttgaactcta actacaccga cgaagcatac
     1861 aagagaagag cagaagaatt acgtggtaaa gtaaagatag ccatcaaaga tgtcatcgaa
     1921 cctttggacc aattggaatt gattgataac ttgcaaagat tgggtttagc ccatagattt
     1981 gaaaccgaaa tcagaaacat cttgaacaac atctataaca acaataagga ttacaactgg
     2041 agaaaggaaa atttgtacgc tacttccttg gaattcagat tgttaagaca acacggttac
     2101 ccagtcagtc aagaagtttt taacggtttc aaggatgacc aaggtggttt tatttgtgat
     2161 gacttcaagg gtatattgtc cttgcatgaa gcttcttact actcattgga aggtgaaagt
     2221 ataatggaag aagcatggca attcacttcc aaacacttga aggaagttat gataagtaaa
     2281 aatatggaag aagatgtttt cgtagctgaa caagcaaaga gagccttgga attaccattg
     2341 cattggaaag ttcctatgtt ggaagcaaga tggttcatcc atatatacga aagaagagaa
     2401 gataagaacc acttgttgtt ggaattggca aagatggaat tcaatacatt acaagccatc
     2461 tatcaagaag aattgaagga aatctctggt tggtggaagg ataccggttt gggtgaaaag
     2521 ttgtcattcg ctagaaatag attggtcgca tctttcttat ggtcaatggg tattgccttt
     2581 gaacctcaat tcgcttactg tagaagagtt ttgacaatct ctattgcatt gatcaccgtt
     2641 atagatgaca tctatgacgt atacggtact ttggatgaat tggaaatctt tacagacgct
     2701 gttgaaagat gggatatcaa ctatgcatta aagcatttgc caggttacat gaagatgtgc
     2761 tttttagcct tgtacaactt cgtcaacgaa ttcgcttact acgttttgaa gcaacaagat
     2821 ttcgacttat tgttatctat taaaaacgct tggttgggtt tgatacaagc ctatttggta
     2881 gaggctaagt ggtaccattc taagtacaca cctaagttgg aagaatactt ggaaaacggt
     2941 ttagtctcaa tcactggtcc attgatcatc acaatctcct atttgagtgg tactaaccct
     3001 atcattaaaa aggaattgga attcttggaa tcaaacccag atatcgttca ctggtcttca
     3061 aaaattttca gattgcaaga tgacttaggt acttccagtg acgaaattca aagaggtgac
     3121 gtacctaaat ctatacaatg ctacatgcat gaaacaggtg catcagaaga agttgccaga
     3181 caacacatta aggatatgat gagacaaatg tggaaaaagg ttaatgctta caccgcagat
     3241 aaagactccc cattgactgg tactacaacc gaattcttgt tgaacttagt tagaatgtct
     3301 catttcatgt atttgcatgg tgacggtcac ggtgtacaaa atcaagaaac cattgatgtc
     3361 ggttttactt tgttattcca acctattcca ttagaagata aacacatggc attcaccgca
     3421 tcaccaggta caaagggtta atagctcgag atagattaat ttaaacagta tatgtacagt
     3481 tttatatata tatatatata tatatacata tataaagaaa cctgtgcgtt ttttgtattt
     3541 tcaaatacat ttagttttgg cgccacttct ataaaaggtc tcatgatatt gttactctag
     3601 agatcccaat acaacagatc acgtgatctt ttgtaagatg aagttgaagt gagtgttgca
     3661 ccgtgccaat gcaggtggct attagattaa atatgtgatt tgttctatta agtttcctgt
     3721 ataattaatg gggagcgctg attctctttt ggtacgcttc ccatccagca tttctgtatc
     3781 tttcaccttc aaccttagga tctctaccct tggcgaaaag tcctctgcca acaatgatga
     3841 tatctgatcc accacttaca acttcgtcga cggttctgta ctgctgaccc aatgcatcgc
     3901 ctttgtcgtc taaacctaca cctggggtca tgattagcca atcaaaccct tcttctcttc
     3961 ctcccatatc gttctgagca atgaacccaa taacgaaatc tttatcactc tttgcaatat
     4021 caacggtacc cttagtatat tcaccgtgtg ctagagaacc cttggaagac aattcagcaa
     4081 gcatcaataa tccccttggt tctttggtga cctcttgcgc accttgtttc aagccagcaa
     4141 caataccagc accagtaacc ccgtgggcgt tggtgatatc agaccattct gcgatacggt
     4201 aaacgcccga tgtatattgt aatttgactg tgttaccgat atcggcgaat tttctgtcct
     4261 caaatatcaa gaacttgtat ttctctgcca atgctttcaa tggaacgaca gtaccctcat
     4321 aactgaaatc atccaagata tcaacgtgtg ttttcaaaag gcaaatgtat ggacccaacg
     4381 tttcaacaag tttcaatagc tcatcagtcg aacgaacgtc aagagaagca cacaaattgg
     4441 tcttcttttc atccattaaa cgtaaaagtt tcgatgcaac cggacttgca tgagtctcag
     4501 ctctactggt atatgatttt gtggacatgt tttatcttcg tttcctgcag gtttttgttc
     4561 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacca catatgcgta
     4621 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgctcg gagattaccg
     4681 aatcaaaaaa atttaaatga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4741 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4801 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4861 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     4921 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4981 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5041 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5101 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5161 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5221 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5281 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5341 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5401 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5461 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5521 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5581 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5641 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5701 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5761 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5821 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5881 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5941 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6001 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6061 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6121 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6181 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6241 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6301 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6361 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6421 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     6481 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6541 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6601 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6661 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6721 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6781 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6841 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6901 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6961 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7021 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7081 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7141 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7201 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     7261 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     7321 ccatgattac gccaagcttg catgcattta aatcattacg accgagattc ccgggtaata
     7381 actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata
     7441 atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc
     7501 tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct actag
//
