LOCUS       sequence-168930-        7428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive TEF1 promoter mediated expression of the UBiC and ARO4*
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  AUTHORS   Williams TC, Peng B, Vickers CE, Nielsen LK
  TITLE     The Saccharomyces cerevisiae pheromone-response is a metabolically 
            active stationary phase for bio-production.
  JOURNAL   Metab Eng Commun. 2016 May 11;3:142-152. doi: 
            10.1016/j.meteno.2016.05.001. eCollection 2016 Dec.
  PUBMED    29468120
REFERENCE   2  (bases 1 to 7428)
  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 
            Commun."; date: "2016-05-11"; volume: "3:142-152. doi"; pages: " 
            10.1016/j.meteno.2016.05.001. eCollection 2016 Dec"
COMMENT     Sequence Label: PHBA02
FEATURES             Location/Qualifiers
     source          1..7428
                     /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"
     promoter        2038..2444
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2962..2978)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3002..3251
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        3294..3682
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(4815..4831)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4857..5104
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5168..5188)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5168..5186)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5207..5223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5207..5223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5220..5242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5231..5247
                     /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(5255..5261,5262..5279,5280..5285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5438..5455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5609..6197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5689..5708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6368..7159,7160..7228))
                     /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     6991..7010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7229..7333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7401..7419
                     /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 cgaggtcgac ctcgaggcac
     2041 acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc
     2101 gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct
     2161 tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc
     2221 tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg
     2281 aaaatttttt tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat
     2341 aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta caactttttt
     2401 tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttaa ttacaaatgt
     2461 cccatccagc tttgacacaa ttgagagctt tgagatactg caaagaaatt ccagcattgg
     2521 atccacaatt attggactgg ttgttgttgg aagattccat gactaagaga ttcgaacaac
     2581 aaggtaagac cgtttccgtt accatgatta gagaaggttt cgtcgaacaa aacgaaatcc
     2641 cagaagaatt gcctttgttg ccaaaagaat ctagatactg gttgagagaa attttgttgt
     2701 gcgctgatgg tgaaccttgg ttggctggta gaactgttgt tccagtttct actttgtctg
     2761 gtccagaatt ggcattgcaa aaattgggta aaactccatt gggtagatac ttgttcactt
     2821 cttctacctt gaccagagac ttcattgaaa ttggtagaga tgctggtttg tggggtagaa
     2881 gatcaagatt gagattgtct ggtaaacctt tgttgttgac cgaattattc ttgccagctt
     2941 ccccattata ctgacccggg ggatccacta gttctagaag gccccttttc ctttgtcgat
     3001 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     3061 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     3121 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaacgcg
     3181 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     3241 ctttaatttg caagcttcgc agtttacact ctcatcgtcg gaattcgctt caaaatgttt
     3301 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
     3361 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
     3421 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
     3481 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt tttgattttt
     3541 ttctctttcg atgacctccc attgatattt aagttaataa acggtcttca atttctcaag
     3601 tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc attagaaaga
     3661 aagcatagca atctaatcta agttttaatt acaaatgagt gaatctccaa tgttcgctgc
     3721 caacggcatg ccaaaggtaa atcaaggtgc tgaagaagat gtcagaattt taggttacga
     3781 cccattagct tctccagctc tccttcaagt gcaaatccca gccacaccaa cttctttgga
     3841 aactgccaag agaggtagaa gagaagctat agatattatt accggtaaag acgacagagt
     3901 tcttgtcatt gtcggtcctt gttccatcca tgatctagaa gccgctcaag aatacgcttt
     3961 gagattaaag aaattgtcag atgaattaaa aggtgattta tccatcatta tgagagcata
     4021 cttggagaag ccaagaacaa ccgtcggctg gaaaggtcta attaatgacc ctgatgttaa
     4081 caacactttc aacatcaaca agggtttgca atccgctaga caattgtttg tcaacttgac
     4141 aaatatcggt ttgccaattg gttctgaaat gcttgatacc atttctccta aatacttggc
     4201 tgatttggtc tccttcggtg ccattggtgc cagaaccacc gaatctcaac tgcacagaga
     4261 attggcctcc ggtttgtctt tcccagttgg tttcaagaac ggtaccgatg gtaccttaaa
     4321 tgttgctgtg gatgcttgtc aagccgctgc tcattctcac catttcatgg gtgttactaa
     4381 gcatggtgtt gctgctatca ccactactaa gggtaacgaa cactgcttcg ttattctaag
     4441 aggtggtaaa aagggtacca actacgacgc taagtccgtt gcagaagcta aggctcaatt
     4501 gcctgccggt tccaacggtc taatgattga ctactctcac ggtaactcca ataaggattt
     4561 cagaaaccaa ccaaaggtca atgacgttgt ttgtgagcaa atcgctaacg gtgaaaacgc
     4621 cattaccggt gtcatgattg aatcaaacat caacgaaggt aaccaaggca tcccagccga
     4681 aggtaaagcc ggcttgaaat atggtgtttc catcactgat gcttgtatag gttgggaaac
     4741 tactgaagac gtcttgagga aattggctgc tgctgtcaga caaagaagag aagttaacaa
     4801 gaaatagccc gggggatcca ctagttctag aacaggcccc ttttcctttg tcgatatcat
     4861 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
     4921 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
     4981 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaac gcgtgtacgc
     5041 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat
     5101 ttgcaagctt cgcagtttac actctcatcg tgcggccgcc accgcggtgg agctccagct
     5161 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     5221 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat aggagccgga agcataaagt
     5281 gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt aattgcgttg cgctcactgc
     5341 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5401 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5461 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5521 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5641 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5701 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5761 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5821 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5881 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5941 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6001 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6061 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7381 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
