LOCUS       sequence-168928-        6439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aromatic amino acid inducible expression of alpha pheromone in yeast
            from the mfalpha1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6439)
  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 6439)
  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: pTCW009
FEATURES             Location/Qualifiers
     source          1..6439
                     /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"
     CDS             join(2813..2869,2870..3067,3068..3079)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYLDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKREAEA"
     primer_bind     complement(3612..3628)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3612..3628)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3665..3685)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3665..3683)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3704..3720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3704..3720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3717..3739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3728..3744
                     /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(3752..3758,3759..3776,3777..3782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3935..3952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4106..4694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4186..4205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4865..5656,5657..5725))
                     /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     5488..5507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5726..5830)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5867..6370
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6412..6430
                     /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 ctgcaccgcc 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 aaaaaaaaag cggtaactcg cggaagcatt
     2641 gccgatgctt accgagattt gccgcggata accgaaccat cattggggta ggaaacatta
     2701 taaatatttg atatatataa aggaaaaagc ttgatttggt cttgtaatat cccagtgtaa
     2761 acataaaaat acacacatac cacaattaca ctctctcatc gactcagaat tcatgagatt
     2821 tccttcaatt tttactgcag ttttattcgc agcatcctcc gcattagctg ctccagtcaa
     2881 cactacaaca gaagatgaaa cggcacaaat tccggctgaa gctgtcatcg gttacttaga
     2941 tttagaaggg gatttcgatg ttgctgtttt gccattttcc aacagcacaa ataacgggtt
     3001 attgtttata aatactacta ttgccagcat tgctgctaaa gaagaagggg tatctttgga
     3061 taaaagagag gctgaagctt ggcattggtt gcaactaaaa cctggccaac caatgtacaa
     3121 gagagaagcc gaagctgaag cttggcattg gctgcaacta aagcctggcc aaccaatgta
     3181 caaaagagaa gccgacgctg aagcttggca ttggctgcaa ctaaagcctg gccaaccaat
     3241 gtacaaaaga gaagccgacg ctgaagcttg gcattggttg cagttaaaac ccggccaacc
     3301 aatgtactaa gcccgactga taacaacagt gtagatgtaa caaagtcgac tttgttccca
     3361 ctgtactttt agctcgtaca aaatacaata tacttttcat ttctccgtaa acaacatgtt
     3421 ttcccatgta atatcctttt ctatttttcg ttccgttacc aactttacac atactttata
     3481 tagctattca cttctataca ctaaaaaact aagacaattt taattttgct gcctgccata
     3541 tttcaatttg ttataaattc ctataattta tcctattagt agctaaaaaa agatgaatgt
     3601 gaatcgaatc ggatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt
     3661 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3721 tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc ataaagtgta
     3781 aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc tcactgcccg
     3841 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3901 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5821 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc cttttcatca cgtgctataa
     5881 aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa
     5941 aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc
     6001 gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt
     6061 ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat
     6121 cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa
     6181 gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac
     6241 tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac
     6301 acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag
     6361 gcaagcgatc cgtcctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6421 atcacgaggc cctttcgtc
//
