LOCUS       sequence-168931-        7582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  UBiC and feedback resistant ARO4 expression from pheromone inducible
            FUS1J2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  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 7582)
  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: pTCW015
FEATURES             Location/Qualifiers
     source          1..7582
                     /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"
     primer_bind     complement(3037..3053)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3077..3326
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4990..5006)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5032..5279
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5322..5342)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5322..5340)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5361..5377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5361..5377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5374..5396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5385..5401
                     /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(5409..5415,5416..5433,5434..5439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5592..5609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5763..6351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5843..5862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6522..7313,7314..7382))
                     /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     7145..7164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7383..7487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7555..7573
                     /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 cgagccctcc ttcaattttt
     2041 ctggcaactt ttctcaacag aacaataacg gcaaccagaa ccgctactga acgatgattc
     2101 agttcgcctt ctatcctttg ttttcgtatt tgtttatata tataacttta ttttttttta
     2161 ttaattgggc tgcaagacaa ttttgttgtc agtgatgcct caatccttct tttgcttcca
     2221 tatttaccat gtggaccctt tcaaaacaga gttgtatctc tgcaggatgc cctttttgac
     2281 gtattgaatg gcataattgc actgtcactt ttcgagctgt cccattttgg tgcgatgatg
     2341 gaacaaacat gaaacgtctg taatttgaaa caaataacgt aattctcggg attggtttta
     2401 tttaaatgac aatgtaagag tggctttgta aggtatgtgt tgctcttaaa atatttggat
     2461 acgacttccg ttatcttttt tcctttaaga gcaggatata agccaacaag tttctgaaaa
     2521 tcaaagaatt catgtcccat ccagctttga cacaattgag agctttgaga tactgcaaag
     2581 aaattccagc attggatcca caattattgg actggttgtt gttggaagat tccatgacta
     2641 agagattcga acaacaaggt aagaccgttt ccgttaccat gattagagaa ggtttcgtcg
     2701 aacaaaacga aatcccagaa gaattgcctt tgttgccaaa agaatctaga tactggttga
     2761 gagaaatttt gttgtgcgct gatggtgaac cttggttggc tggtagaact gttgttccag
     2821 tttctacttt gtctggtcca gaattggcat tgcaaaaatt gggtaaaact ccattgggta
     2881 gatacttgtt cacttcttct accttgacca gagacttcat tgaaattggt agagatgctg
     2941 gtttgtgggg tagaagatca agattgagat tgtctggtaa acctttgttg ttgaccgaat
     3001 tattcttgcc agcttcccca ttatactgac ccgggggatc cactagttct agaaggcccc
     3061 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     3121 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     3181 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     3241 ttctgtacaa acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     3301 tgggacgctc gaaggcttta atttgcaagc ttcgcagttt acactctcat cgtcgagtcg
     3361 agccctcctt caatttttct ggcaactttt ctcaacagaa caataacggc aaccagaacc
     3421 gctactgaac gatgattcag ttcgccttct atcctttgtt ttcgtatttg tttatatata
     3481 taactttatt tttttttatt aattgggctg caagacaatt ttgttgtcag tgatgcctca
     3541 atccttcttt tgcttccata tttaccatgt ggaccctttc aaaacagagt tgtatctctg
     3601 caggatgccc tttttgacgt attgaatggc ataattgcac tgtcactttt cgagctgtcc
     3661 cattttggtg cgatgatgga acaaacatga aacgtctgta atttgaaaca aataacgtaa
     3721 ttctcgggat tggttttatt taaatgacaa tgtaagagtg gctttgtaag gtatgtgttg
     3781 ctcttaaaat atttggatac gacttccgtt atcttttttc ctttaagagc aggatataag
     3841 ccaacaagtt tctgaaaatc aaagaattca tgagtgaatc tccaatgttc gctgccaacg
     3901 gcatgccaaa ggtaaatcaa ggtgctgaag aagatgtcag aattttaggt tacgacccat
     3961 tagcttctcc agctctcctt caagtgcaaa tcccagccac accaacttct ttggaaactg
     4021 ccaagagagg tagaagagaa gctatagata ttattaccgg taaagacgac agagttcttg
     4081 tcattgtcgg tccttgttcc atccatgatc tagaagccgc tcaagaatac gctttgagat
     4141 taaagaaatt gtcagatgaa ttaaaaggtg atttatccat cattatgaga gcatacttgg
     4201 agaagccaag aacaaccgtc ggctggaaag gtctaattaa tgaccctgat gttaacaaca
     4261 ctttcaacat caacaagggt ttgcaatccg ctagacaatt gtttgtcaac ttgacaaata
     4321 tcggtttgcc aattggttct gaaatgcttg ataccatttc tcctaaatac ttggctgatt
     4381 tggtctcctt cggtgccatt ggtgccagaa ccaccgaatc tcaactgcac agagaattgg
     4441 cctccggttt gtctttccca gttggtttca agaacggtac cgatggtacc ttaaatgttg
     4501 ctgtggatgc ttgtcaagcc gctgctcatt ctcaccattt catgggtgtt actaagcatg
     4561 gtgttgctgc tatcaccact actaagggta acgaacactg cttcgttatt ctaagaggtg
     4621 gtaaaaaggg taccaactac gacgctaagt ccgttgcaga agctaaggct caattgcctg
     4681 ccggttccaa cggtctaatg attgactact ctcacggtaa ctccaataag gatttcagaa
     4741 accaaccaaa ggtcaatgac gttgtttgtg agcaaatcgc taacggtgaa aacgccatta
     4801 ccggtgtcat gattgaatca aacatcaacg aaggtaacca aggcatccca gccgaaggta
     4861 aagccggctt gaaatatggt gtttccatca ctgatgcttg tataggttgg gaaactactg
     4921 aagacgtctt gaggaaattg gctgctgctg tcagacaaag aagagaagtt aacaagaaat
     4981 agcccggggg atccactagt tctagaacag gccccttttc ctttgtcgat atcatgtaat
     5041 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     5101 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     5161 gaacgttatt tatatttcaa atttttcttt tttttctgta caaacgcgtg tacgcatgta
     5221 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     5281 agcttcgcag tttacactct catcgtctcc agcttttgtt ccctttagtg agggttaatt
     5341 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5401 attccacaca acataggagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5461 aggtaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5521 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6181 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7501 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     7561 cgtatcacga ggccctttcg tc
//
