LOCUS       sequence-176087-        7373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of prespatane synthase LptTPS-A in S. cerevisiae for 
            sesquiterpene production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7373)
  TITLE     Prespatane synthase yeast expression plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7373)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p426TEF-LptTPS-A
FEATURES             Location/Qualifiers
     source          1..7373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    895..916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(931..936,937..954,955..961)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    969..985
                     /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     974..996
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     993..1009
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     993..1009
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1028..1048
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1030..1048
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1066..1464
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2550..2566)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2551..2567)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2569..2816
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2585..2603)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2834..2853)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2835..2853)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2863..2880)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2863..2879)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2872..2894)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3020..3475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3107..3126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3317..3338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3606..4409)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4410..4625)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4649..4668)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4768..4790
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4828..4846)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      4887..6229
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5219..5266
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        6256..6360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6361..6429,6430..7221)
                     /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(6579..6598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      841 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      901 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      961 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1021 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcatag cttcaaaatg
     1081 tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt
     1141 caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa
     1201 ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc
     1261 gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt tttttgattt
     1321 ttttctcttt cgatgacctc ccattgatat ttaagttaat aaacggtctt caatttctca
     1381 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgc tcattagaaa
     1441 gaaagcatag caatctaatc taagttttct agaactagta tgtcattagc caataacatc
     1501 gctccgacac actcaatgag atcagattca gtcgaagttg gcttcaataa gctaaggttt
     1561 accagtttta cgagtttcgg cgacgagttc ataaacgaac atgaggctcc tgcatttatc
     1621 gagtcagttg cgtggtttca gtccctgaat gcaatagcta ccccacagca tctaaaaata
     1681 gtgaaaaatg cgaccttcga gcgtttggta agtcgtactt tcccctttgc cgatttggcc
     1741 ggagctcgta tcgcaactga ccttatgatt ctgacgtttc tgatcgatga tttatcagat
     1801 gtcgtagaag ctacagacga tacagctatg catgcgatgt cagcggtgga aggacaggta
     1861 acgcatgtat tgcgtggcgg aactcccaga cctggagaac atcctcttgc cgtagcaatg
     1921 cgttcaatcg tcgaccgtgc catgttgact tataacccgg actggattga cttaatgcgt
     1981 aaagaattca tcacatacct tgaaatgaat agactagagc gtataaatag gttagaaggt
     2041 ccaggcttga gttggaccat gttcgagaat acgaggtatt actcatcctg cgtgttgcct
     2101 tttttatacc ttagcgcagg catgggttgc acgggctgtc cttccaccgt cttgagtgta
     2161 ccatttgtca agataatgac tgatctaact gtgaatcatg tggcgtgggt aaatgacatc
     2221 gtgggcgcaa ataaagaaag aaaagaagcc gtcaacaata atattgtttt cgtgattgct
     2281 aatgacaggg gtctgacaat ggcaggagcg gtgaaagatg cggtgaaaag gacaaaccaa
     2341 gaatgtgagg tgttcttaaa tctggagcac cgtcttcacg ccggaggagc agtcgttgac
     2401 ggcgatgact tattcaatta tatagaggtg ctaaagtatt ggatgagagg ctccttagac
     2461 tggcatttcg agtctaaaag atacaaggtc aaagcttcta agtagactag tggatccccc
     2521 gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgagtc atgtaattag
     2581 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     2641 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     2701 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     2761 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc
     2821 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     2881 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2941 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3001 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3061 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3361 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3421 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     3481 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga
     3541 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     3601 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     3661 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     3721 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     3781 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     3841 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     3901 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     3961 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     4021 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     4081 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     4141 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     4201 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     4261 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     4321 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     4381 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     4441 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     4501 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     4561 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     4621 ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4681 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4741 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4801 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4861 tcatgataat aatggtttct tagtatgatc caatatcaaa ggaaatgata gcattgaagg
     4921 atgagactaa tccaattgag gagtggcagc atatagaaca gctaaagggt agtgctgaag
     4981 gaagcatacg ataccccgca tggaatggga taatatcaca ggaggtacta gactaccttt
     5041 catcctacat aaatagacgc atataagtac gcatttaagc ataaacacgc actatgccgt
     5101 tcttctcatg tatatatata tacaggcaac acgcagatat aggtgcgacg tgaacagtga
     5161 gctgtatgtg cgcagctcgc gttgcatttt cggaagcgct cgttttcgga aacgctttga
     5221 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttcagag cgcttttgaa
     5281 aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
     5341 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
     5401 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
     5461 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
     5521 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
     5581 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
     5641 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
     5701 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
     5761 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
     5821 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
     5881 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     5941 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     6001 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     6061 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt
     6121 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     6181 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgttc aggtggcact
     6241 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6301 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6361 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6421 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6481 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6541 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6601 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6661 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6721 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6781 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6841 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6901 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6961 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7021 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7081 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7141 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7201 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7261 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt aga
//
