LOCUS       sequence-159453-        6292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aromatic amino acid inducible expression of alpha pheromone in yeast
            from the mfalpha2 gene .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6292)
  AUTHORS   Williams TC, Nielsen LK, Vickers CE
  TITLE     Engineered quorum sensing using pheromone-mediated cell-to-cell 
            communication in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2013 Mar 15;2(3):136-49. doi: 10.1021/sb300110b. 
            Epub 2013 Jan 7.
  PUBMED    23656437
REFERENCE   2  (bases 1 to 6292)
  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"; doi: "10.1021/sb300110b";
            journalName: "ACS Synth Biol"; date: "2013-03-15- 15"; volume: "2"; 
            issue: "3"; pages: "136-49"
COMMENT     Sequence Label: pTCW010
FEATURES             Location/Qualifiers
     source          1..6292
                     /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"
     primer_bind     complement(3518..3538)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3518..3536)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3557..3573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3557..3573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3570..3592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3581..3597
                     /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(3605..3611,3612..3629,3630..3635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3788..3805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3959..4547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4039..4058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4718..5509,5510..5578))
                     /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     5341..5360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5579..5683)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5720..6223
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6265..6283
                     /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 agctccagct tttgttccct ttagtgaggg ttaattgcgc
     3541 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3601 cacacaacat aggagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt
     3661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4381 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4681 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4801 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4861 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4921 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5161 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5221 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5401 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5581 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5641 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5701 tgccacctgg gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat
     5761 ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg
     5821 ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc
     5881 tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca
     5941 cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat
     6001 ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa
     6061 cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta
     6121 aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa attattccat
     6181 cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtccta agaaaccatt
     6241 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
