LOCUS       sequence-159458-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pheromone mediated alpha pheromone expression in yeast from the 
            mfalpha 2 gene and FUSJ2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  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 6084)
  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: pTCW008
FEATURES             Location/Qualifiers
     source          1..6084
                     /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(3257..3273)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3257..3273)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3310..3330)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3310..3328)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3349..3365)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3349..3365)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3362..3384)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3373..3389
                     /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(3397..3403,3404..3421,3422..3427))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3580..3597)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3751..4339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3831..3850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4510..5301,5302..5370))
                     /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     5133..5152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5371..5475)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5512..6015
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6057..6075
                     /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 ctcgagccct
     2101 ccttcaattt ttctggcaac ttttctcaac agaacaataa cggcaaccag aaccgctact
     2161 gaacgatgat tcagttcgcc ttctatcctt tgttttcgta tttgtttata tatataactt
     2221 tatttttttt tattaattgg gctgcaagac aattttgttg tcagtgatgc ctcaatcctt
     2281 cttttgcttc catatttacc atgtggaccc tttcaaaaca gagttgtatc tctgcaggat
     2341 gccctttttg acgtattgaa tggcataatt gcactgtcac ttttcgagct gtcccatttt
     2401 ggtgcgatga tggaacaaac atgaaacgtc tgtaatttga aacaaataac gtaattctcg
     2461 ggattggttt tatttaaatg acaatgtaag agtggctttg taaggtatgt gttgctctta
     2521 aaatatttgg atacgacttc cgttatcttt tttcctttaa gagcaggata taagccaaca
     2581 agtttctgaa aatcaaagaa ttcatgaaat tcatttctac ctttctcact tttattttag
     2641 cggccgtttc tgtcactgct agttccgatg aagatatcgc tcaggtgcca gccgaggcca
     2701 ttattggata cttggatttc ggaggtgatc atgacatagc ttttttacca ttcagtaacg
     2761 ctaccgccag tgggctattg tttatcaaca ccactattgc tgaggcggct gaaaaagagc
     2821 aaaacaccac tttggcgaaa agagaggctg ttgccgacgc ttggcactgg ttaaatttga
     2881 gaccaggcca accaatgtac aagagagagg ccaacgctga tgcttggcac tggttgcaac
     2941 tcaagccagg ccaaccaatg tactgaaaaa tgaccctaaa ctacttctaa accctctcga
     3001 tttctttcac gttcatacaa cacctagttt tatttatttt cttttcaatc tgagtagttg
     3061 agttttcgat cactcacata gaactatttt ttgccattta aataaagtat tctctcaaat
     3121 gatgcgatac tataatactc tttgccatat attacattca ttcataaata ggctatgttt
     3181 ctatatccgt ttccgattct gtctgcaagc aaggttccct atcattaccg gattgttcac
     3241 tatggttgga gctctggatc cactagttct agagcggccg ccaccgcggt ggagctccag
     3301 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     3361 tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa
     3421 gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact
     3481 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3541 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3601 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3661 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3721 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3781 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3841 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3901 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3961 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4021 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4081 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4141 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4201 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4261 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4321 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4381 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4441 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4501 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4561 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4621 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4681 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4741 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4801 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4861 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4921 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4981 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5041 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5101 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5161 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5221 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5281 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5341 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5401 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5461 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc
     5521 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     5581 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     5641 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     5701 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     5761 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     5821 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     5881 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     5941 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     6001 tacaggcaag cgatccgtcc taagaaacca ttattatcat gacattaacc tataaaaata
     6061 ggcgtatcac gaggcccttt cgtc
//
