LOCUS       sequence-168904-        6535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pheromone mediated destablised-GFP expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6535)
  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 6535)
  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: pTCW002
FEATURES             Location/Qualifiers
     source          1..6535
                     /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        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1936..1958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1950..1967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1977..1996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2756..3469
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4027..4214
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4082..4106)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4222..4238)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4222..4238)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4275..4295)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4275..4293)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4314..4330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4314..4330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4327..4349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4338..4354
                     /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(4362..4368,4369..4386,4387..4392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4545..4562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4716..5304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4796..4815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5475..6266,6267..6335))
                     /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     6098..6117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6336..6440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6508..6526
                     /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 tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgagatcaac aacagggtca
     2041 gcagcagcaa caaggccact ccagttcatt ctcagctttg gcttccatgg caagttccta
     2101 cctgggcaat aactccaatt caaattcgag ttatgggggc cagcaacagg ctaatgagta
     2161 tggtagaccg caacagaatg gtcaacagca atccaatgag tacggaagac cgcaatacgg
     2221 cggaaaccag aactccaatg gacagcacga atccttcaat ttttctggca acttttctca
     2281 acagaacaat aacggcaacc agagccgcta ctgaacgatg attcagttcg ccttctatcc
     2341 tttgtttacg tatttgttta tatatataac tttatttttt tttattaatt gggctgcaag
     2401 acaattttgt tgtcagtgat gcctcaatcc ttcttttgct tccatattta ccatgtggac
     2461 cctttcaaaa cagagttgta tctctgcagg atgccctttt tgacgtattg aatggcataa
     2521 ttgcactgtc acttttcgcg ctgtctcatt ttggtgcgat gatgaaacaa acatgaaacg
     2581 tctgtaattt gaaacaaata acgtaattct cgggattggt tttatttaaa tgacaatgta
     2641 agagtggctt tgtaaggtat gtgttgctct taaaatattt ggatacgaca tcctttatct
     2701 tttttccttt aagagcagga tataagccat caagtttctg aaaatcaaag aattcatgtc
     2761 taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt
     2821 taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt
     2881 gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac
     2941 tactttcggt tatggtgttc aatgttttgc gagataccca gatcatatga aacaacatga
     3001 ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga
     3061 tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag
     3121 aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga
     3181 atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa
     3241 agttaacttc aaaattagac acaacattga agatggttct gttcaattag ctgaccatta
     3301 tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc
     3361 cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga
     3421 atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaag catccaactt
     3481 gaacatttcg agaaagctta ccatatcaac cccatcatgc tctttcgaaa attcaaatag
     3541 cacatccatt ccttcgcccg cttcctcatc tcaaagccac actccaatga gaaacatgag
     3601 ctcactctct gataacagcg ttttcagccg gaatatggaa caatcatcac caatcactcc
     3661 aagtatgtac caatttggtc agcagcagtc aaacagtata tgtggtagca ccgttagtgt
     3721 gaatagtctg gtgaatacaa ataacaaaca aaggatctac gaacaaatca cgggtcctaa
     3781 cagcaataac gcaaccaatg attatattga tttgctaaac ctaaatgagt ctaacaagga
     3841 aaaccaaaat cccgcaacgg cgcattacct caatgggggc ccacccaaga caagcttcat
     3901 taaccatgga atgttcccct cgccaactgg gaccataaat agcggtaaat ctagcagtgc
     3961 ctcatcttta atttcttttg gtatgggcaa tacccaagta atatagggcg cgccacttct
     4021 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     4081 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     4141 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
     4201 cacacctcta ccggcagatc ggatccacta gttctagagc ggccgccacc gcggtggagc
     4261 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
     4381 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
     4441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6481 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
