LOCUS       sequence-159463-        7693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aromatic amino acid inducible expression of alpha pheromone, and 
            pheromone inducible exression of an ARO7 RNA interference hairpin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7693)
  AUTHORS   Williams TC, Averesch NJH, Winter G, Plan MR, Vickers CE, Nielsen 
            LK, Kromer JO
  TITLE     Quorum-sensing linked RNA interference for dynamic metabolic pathway
            control in Saccharomyces cerevisiae.
  JOURNAL   Metab Eng. 2015 May;29:124-134. doi: 10.1016/j.ymben.2015.03.008. 
            Epub 2015 Mar 16.
  PUBMED    25792511
REFERENCE   2  (bases 1 to 7693)
  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: "2015-05"; volume: "29:124-134. doi"; pages: " 
            10.1016/j.ymben.2015.03.008. Epub 2015 Mar 16"
COMMENT     Sequence Label: pTCW018
FEATURES             Location/Qualifiers
     source          1..7693
                     /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"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1163
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1560..1581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1772..1791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2049..2068
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3465..3481)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3465..3481)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4626..4875
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4642..4660)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4919..4939)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4919..4937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4958..4974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4958..4974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4971..4993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4982..4998
                     /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(5006..5012,5013..5030,5031..5036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5189..5206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5360..5948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5440..5459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6119..6910,6911..6979))
                     /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     6742..6761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6980..7084)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7121..7624
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7666..7684
                     /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 tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgagttgc
     2101 cgcgtggaga catctgtcca atttgcccct ggctctctgt aaatacccat atatgaacgt
     2161 tatatatatt gtatattaat ggcgtatatg tgcccatggg aagtcatagt aatagatcta
     2221 ggcacgtggc tgcagtgact ccgcgcaggg ctgtaccgcc aaggtagata agagagcgga
     2281 gcacgtggga aaagagagat acggtatgtt acaacagtca aagtaatgaa ctttcgccca
     2341 cgtgcgtaca tcatttacta tctcaggccg ctgcgtttgg aactttacgg cgttgtagtt
     2401 gtaattgcca taactgccat gcatttatgt atatatattt tcgtacgtaa tgaacatggt
     2461 tatgttatat attgtttgca atcagagaga agttgaagag gagaaaaaaa aaaagagaga
     2521 gccgtaccgc aataaaatgg acccttgtcg gtcacgtgag caaaaatact attgccgccg
     2581 acggccgtag ataataacaa aggaataaaa aaaaaaaagc ggtaactcgc ggaagcattg
     2641 ccgatgctta ccgagatttg ccgcggataa ccgaaccatc attggggtag gaaacattat
     2701 aaatatttga tatatataaa ggaaaaagct tgatttggtc ttgtaatatc ccagtgtaaa
     2761 cataaaaata cacacatacc acaattacac tctctcatcg actcagaatt catgaaattc
     2821 atttctacct ttctcacttt tattttagcg gccgtttctg tcactgctag ttccgatgaa
     2881 gatatcgctc aggtgccagc cgaggccatt attggatact tggatttcgg aggtgatcat
     2941 gacatagctt ttttaccatt cagtaacgct accgccagtg ggctattgtt tatcaacacc
     3001 actattgctg aggcggctga aaaagagcaa aacaccactt tggcgaaaag agaggctgtt
     3061 gccgacgctt ggcactggtt aaatttgaga ccaggccaac caatgtacaa gagagaggcc
     3121 aacgctgatg cttggcactg gttgcaactc aagccaggcc aaccaatgta ctgaaaaatg
     3181 accctaaact acttctaaac cctctcgatt tctttcacgt tcatacaaca cctagtttta
     3241 tttattttct tttcaatctg agtagttgag ttttcgatca ctcacataga actatttttt
     3301 gccatttaaa taaagtattc tctcaaatga tgcgatacta taatactctt tgccatatat
     3361 tacattcatt cataaatagg ctatgtttct atatccgttt ccgattctgt ctgcaagcaa
     3421 ggttccctat cattaccgga ttgttcacta tggttggagc tctggatcca ctagttctag
     3481 agcggccgcc accgcggtgg aggctcccct ccttcaattt ttctggcaac ttttctcaac
     3541 agaacaataa cggcaaccag aaccgctact gaacgatgat tcagttcgcc ttctatcctt
     3601 tgttttcgta tttgtttata tatataactt tatttttttt tattaattgg gctgcaagac
     3661 aattttgttg tcagtgatgc ctcaatcctt cttttgcttc catatttacc atgtggaccc
     3721 tttcaaaaca gagttgtatc tctgcaggat gccctttttg acgtattgaa tggcataatt
     3781 gcactgtcac ttttcgagct gtcccatttt ggtgcgatga tggaacaaac atgaaacgtc
     3841 tgtaatttga aacaaataac gtaattctcg ggattggttt tatttaaatg acaatgtaag
     3901 agtggctttg taaggtatgt gttgctctta aaatatttgg atacgacttc cgttatcttt
     3961 tttcctttaa gagcaggata taagccaaca agtttctgaa aatcaaaatc gattgaaact
     4021 cccttctttc ctgacaagat tcagaaatca ttcttaccga gcattaacta cccacaaatt
     4081 ttggcgcctt atgccccaga agttaattac aatgataaaa taaaaaaagt ttatattgaa
     4141 aagattatac cattaatttc gaaaagagat ggtgatgata agaataactt cggttctgtt
     4201 gccactagag atatagaatg tttgcaaagc ttgagtagga gaatccactt tggcaagttt
     4261 gttctcgaga aaatcaggtg tgttggaact tttttcaaac cttactaaac attgaaacta
     4321 attggtaaag atagaggaat tcaacaaact tgccaaagtg gattctccta ctcaagcttt
     4381 gcaaacattc tatatctcta gtggcaacag aaccgaagtt attcttatca tcaccatctc
     4441 ttttcgaaat taatggtata atcttttcaa tataaacttt ttttatttta tcattgtaat
     4501 taacttctgg ggcataaggc gccaaaattt gtgggtagtt aatgctcggt aagaatgatt
     4561 tctgaatctt gtcaggaaag aagggagttt cacccgggcc ggaggcccct tttcctttgt
     4621 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     4681 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     4741 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     4801 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     4861 aaggctttaa tttgcaagct tcgcagttta cactctcatc gtcctccagc ttttgttccc
     4921 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4981 attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct
     5041 ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc
     5101 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5161 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5221 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5281 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5341 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5401 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5461 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5521 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5581 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5641 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5701 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5761 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5821 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5881 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6181 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6241 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6301 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6361 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6421 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6481 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6541 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6601 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6661 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6721 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6781 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6841 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6901 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6961 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7081 cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa
     7141 ttataattta aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa
     7201 ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac
     7261 aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc
     7321 ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa
     7381 tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc
     7441 tttttgttga aattttttaa acctttgttt attttttttt cttcattccg taactcttct
     7501 accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag
     7561 taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc
     7621 gatccgtcct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7681 aggccctttc gtc
//
